Movies

The following movies require Flash Player. If you don't have this on your machine you can download Flash Player here. If you would like to use my movies for teaching or other purposes please do not hesitate to email a request on G.Steinberg@exeter.ac.uk.

1. In vitro motility assays

View the 'Bead movement by fungal kinesin' movie

a. Bead movement by fungal kinesin
Purified kinesin-1 from Neurospora crassa moves latex beads along pig brain microtubules. The movie runs at real time, the velocity of this movement is approx 1 micrometer per second. Reference: Steinberg and Schliwa, 1995, Mol Biol Cell, 6, 1605.

View the 'Bead movement by fungal kinesin' movie
(832Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 708Kb

Download the PC file - 1.9Mb

View the 'Microtubule gliding over fungal kinesin' movie

b. Microtubule gliding over fungal kinesin
Pig brain microtubules slide over glass-attached kinesin-1 from Neurospora crassa. The movie runs at real time, the velocity of this movement is approx 2-3 micrometer per second. Reference: Steinberg and Schliwa, 1995, Mol Biol Cell, 6, 1605.

View the 'Microtubule gliding over fungal kinesin' movie
(760Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.6Mb

Download the PC file - 1.8Mb

View the 'Axoneme moving over fungal kinesin' movie

c. Axoneme moving over fungal kinesin
A Chlamydomonas axoneme (central body of the flagellum) slides over glass-attached kinesin-1 from Neurospora crassa. The movie runs at real time, the velocity of this movement is approx 0.7 micrometer per second. Note that the pointed end represents the minus-ends of the microtubules in the axoneme. Neurospora kinesin moves to plus-ends and pushes the pointed end of the axoneme forward. Reference: Steinberg and Schliwa, 1995, Mol Biol Cell, 6, 1605.

View the 'Axoneme moving over fungal kinesin' movie
(520Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 404Kb

Download the PC file - 512Kb

View the 'Binding of GFP-labelled Myosin-XVII' movie

d. Binding of GFP-labelled Myosin-XVII to rhodamine-labeled and phalloidin-stabilised actin filaments
Time is given in seconds:milliseconds, Bar represents micrometers. Reference: Schuster et al., 2011, EMBO J., 31, 214-327. View a PDF and more movies.

View the 'Binding of GFP-labelled Myosin-XVII' movie
(191Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 287Kb

Download the PC file - 2Mb

2. Membrane trafficking

View the 'Fusion and fission of mitochondria in Neurospora crassa' movie

a. Fusion and fission of mitochondria in Neurospora crassa
Motility of mitochondria in a wall-less N. crassa mutant, defective in β-D-glucan synthase (fz;sg;os-1; FGSC 1118), in phosphate buffered saline. The movie runs at real time, the elapsed time is given in minutes:seconds. Bar represents micrometers. Reference: Steinberg et al., 1993, J. Cell Sci., 106, 555-564. Download a PDF.

View the 'Fusion and fission of mitochondria in Neurospora crassa' movie
(1.4Mb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.6Mb

Download the PC file - 2.1Mb

View the 'Nuclear migration in Neurospora crassa' movie

b. Nuclear migration in Neurospora crassa
Nuclear migration in a wall-less N. crassa mutant, defective in β-D-glucan synthase (fz;sg;os-1; FGSC 1118), in phosphate buffered saline. The movie runs at real time, the elapsed time is given in minutes:seconds. Reference: Steinberg et al., 1993, J. Cell Sci., 106, 555-564. Download a PDF.

View the 'Nuclear migration in Neurospora crassa' movie
(952Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.3Mb

Download the PC file - 1.6Mb

View the 'Neurospora_Spitzenkörper' movie

c. Dynamics of vesicles in the Spitzenkörper of Neurospora crassa
N. crassa hypha that grow through low melt agarose. Membranous vesicles accumulate in the hyphal tip. A pseudo coloured detail is given left. Elapsed time is given in minutes:seconds. Reference: Steinberg 2007, Euk. Cell, 6, 351-360. Download a PDF.

View the 'Dynamics of vesicles in the Spitzenkörper of Neurospora crassa' movie
(1Mb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1Mb

Download the PC file - 1.4Mb

View the 'Motility of a chitosome' movie

d. Motility of a chitosome towards the growth region in an Ustilago maydis hyphal cell
The endogenous copy of a class V chitin synthase was labeled with a triple-GFP tag. After photo-bleaching of the apical region of the cell, motility of single chitosomes towards the apex can be seen. Time is given in seconds:milliseconds. Bar represents micrometers. Reference: Treitschke et al., 2010, Plant Cell, 22, 2476-2494. More movies and download a PDF.

View the 'Motility of a chitosome' movie
(758Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 546Kb

Download the PC file - 649Kb

View the 'Motility of a chitosome' movie

e. Motility of a chitosome along a fluorescently labled microtubule
The endogenous copy of a class V chitin synthase was labeled with a triple-GFP tag, tubulin was fused to a red-fluorescent protein. Time is given in seconds:milliseconds. Bar represents micrometers. Reference: Schuster et al., 2011, EMBO J., 31, 214-327. More movies and download a PDF.

View the 'Motility of a chitosome' movie
(75Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 48Kb

Download the PC file - 83Kb

View the 'Dynamics of early endosomes in hyphal cells' movie

f. Dynamics of early endosomes in hyphal cells
The organelles move rapidly in a bi-directional fashion. Early endosomes were labelled by a fusion of eGFP to the small GTPase Rab5a. The time is given in seconds:milliseconds; the bar represents 10 micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of early endosomes in hyphal cells' movie
(868Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.34Mb

Download the PC file - 1.57Mb

View the 'Retrograde motility of early endosomes' movie

g. Retrograde motility of early endosomes that carry a photo-activatable GFP-Rab5a fusion protein
The fluorescent fusion protein becomes visible after a laser pulse (yellow arrowhead). The organelles show retrograde motility. Time is given in seconds:milliseconds. Bar represents micrometers. Reference: Schuster et al., 2011, Mol Biol Cell, 22, 3645-3657. More movies and download a PDF.

View the 'Retrograde motility of early endosomes' movie
(674Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 418Kb

Download the PC file - 620Kb

View the 'Retrograde motility of early endosomes' movie

h. Retrograde motility of early endosomes that carry a photo-activatable GFP-Rab5a fusion protein
The fluorescent fusion protein becomes visible after a laser pulse (yellow arrowhead). The organelles show extended retrograde motility. Time is given in seconds:milliseconds. Bar represents micrometers. Reference: Schuster et al., 2011, Mol Biol Cell, 22, 3645-3657. More movies and download a PDF.

View the 'Retrograde motility of early endosomes' movie
(2.6Mb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.89Mb

Download the PC file - 2.39Mb

View the 'Motility of early endosomes' movie

i. Motility of early endosomes along microtubules in a hyphal cell of Ustilago maydis
The organelles move rapidly in a bi-directional fashion along microtubules. Early endosomes were labelled by a fusion of eGFP to the small GTPase Rab5a. Microtubules were visualised by monomeric red fluorescent protein fused to the alpha-tubulin gene tub1. Time is given in seconds:milliseconds. Bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Motility of early endosomes' movie
(539Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 410Kb

Download the PC file - 334Kb

View the 'Behaviour of an early endosome' movie

k. Behaviour of an early endosome at the end of the microtubule track
Dynein (labelled with a triple-red fluorescent protein tag) concentrates at the microtubule plus-end where it forms a "loading zone" and binds to the arriving organelle to take it back towards the cell center. Time is given in seconds:milliseconds. Bar represents micrometers. Reference: Schuster et al., 2011, EMBO J., 30, 652-664. Download a PDF.

View the 'Behaviour of an early endosome' movie
(765Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 491Kb

Download the PC file - 748Kb

3. In vivo motility of molecular motors

View the 'Dynamics of dynein in hyphal cells' movie

a. Dynamics of dynein in hyphal cells
Dynein moves bi-directionally and concentrates at the microtubule plus-ends near the hyphal tip. Note that anterograde motility of dynein in fungi is most likely due to the activity of kinesin-1. Dynein was labelled by fusing a triple GFP tag to the N-terminus of the dyn2 dynein heavy chain gene. The construct is integrated in the native locus; therefore native levels of the motors are shown. Time is given in minutes:seconds. Bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of dynein in hyphal cells' movie
(491Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 756Kb

Download the PC file - 959Kb

View the 'Motility of the dynein motor complex' movie

b. Motility of the dynein motor complex
A fusion protein of a triple-GFP tag N-terminally fused to the endogenous dynein heavy chain gene dyn2 displays bi-directional motility. Most of the dynein concentrates at the MT plus-ends near the hyphal tip. Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Schuster et al., 2011, EMBO J., 30, 652-664. Download a PDF.

View the 'Motility of the dynein motor complex' movie
(487Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 443Kb

Download the PC file - 1.39Mb

View the 'Release of photo-activated dynein from apical MT plus-ends' movie

c. Release of photo-activated dynein from apical MT plus-ends
A triple-tag of photo-activatable GFP was fused to the endogenous copy of the dynein heavy chain. After activation by a 405 nm pulse, apical dynein becomes visible that leaves the tip for retrograde movement. Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Schuster et al., 2011, EMBO J., 30, 652-664. Download a PDF.

View the 'Release of photo-activated dynein from apical MT plus-ends' movie
(520Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 465Kb

Download the PC file - 705Kb

View the 'Dynamics of kinesin-3 in hyphal cells' movie

d. Dynamics of kinesin-3 in hyphal cells
Kinesin-3-GFP continuously move in a bi-directional fashion. Note that the microtubule plus-ends are concentrated at the hyphal tip and that transport towards the hyphal tips is driven by kinesin-3. In contrast, retrograde transport back to the cell centre is mediated by dynein. Kinesin-3 was labelled by fusing eGFP to its C-terminal. The construct is integrated in the native locus; therefore native levels of the motors are shown. The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of kinesin-3 in hyphal cells' movie
(1Mb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 691Kb

Download the PC file - 2.07Mb

View the 'Bi-directional Kin3-GFP motility' movie

e. Bi-directional Kin3-GFP motility
A GFP tag was fused to the C-terminus of kin3, the kinesin-3 in U. maydis. Single signals show processive and b-directional motility (red arrowhead). Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Schuster et al., 2011, PNAS, 108, 3618-3623. Download a PDF.

View the 'Bi-directional Kin3-GFP motility' movie
(243Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 204Kb

Download the PC file - 625Kb

View the 'Dynamics of myosin-5 in a hyphal cell of Ustilgo maydis' movie

f. Dynamics of myosin-5 in a hyphal cell of Ustilgo maydis
Pre-treatment with a 405 nm laser photo-bleaches the cell (red box), allowing the observation of myosin-5 motors moving to the hyphal tip. Myosin-5 was labelled by fusing a triple GFP tag to the N-terminus of the myo5 gene. The construct is integrated in the native locus; therefore native levels of the motor are shown. The time is given in seconds:milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of myosin-5 in a hyphal cell of Ustilgo maydis' movie
(239Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 263Kb

Download the PC file - 1.2Mb

View the 'Motility of fluorescent myosin-5 in a yeast-like cell of Ustilgo maydis' movie

g. Motility of fluorescent myosin-5 in a yeast-like cell of Ustilgo maydis
Myosin-5 was labelled by fusing a triple GFP tag to the N-terminus of the myo5 gene. The construct is integrated in the native locus; therefore native levels of the motor are shown. Note that all signals move at the cell periphery and towards the accumulation at the tip. The time is given in seconds:milliseconds; the bar represents micrometers. Reference: Schuster et al., 2011, EMBO J., 31, 214-327. View more movies and download a PDF.

View the 'Motility of fluorescent myosin-5 in a yeast-like cell of Ustilgo maydis' movie
(429Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 233Kb

Download the PC file - 363Kb

View the 'Co-localization of dynein and early endosomes

h. Co-localization of dynein and early endosomes
A triple-tag of GFP was fused to the endogenous copy of the dynein heavy chain gene dyn2 and the fusion protein was co-expressed with mCherry-Rab5a (red). The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Schuster et al., 2011, PNAS, 108, 3618-3623. Download a PDF.

View the 'Co-localization of dynein and early endosomes' movie
(146Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 224Kb

Download the PC file - 303Kb

View the 'Motility of early endosomes' movie

i. Motility of early endosomes along microtubules in a hyphal cell of Ustilago maydis
A triple-tag of GFP was fused to the endogenous copy of the dynein heavy chain gene dyn2 and the fusion protein was co-expressed with mCherry-Rab5a (red). Several different signals move from the hyphal tip towards the cell center (retrograde): (i) dynein that is not bound to an early endosome (green; 1), (ii) dynein that colocalizes with Rab5-positive early endosomes (green+red=yellow; 2) and (iii) with Rab5-positive early endosomes that move without dynein (red, 3). The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Schuster et al., 2011, Mol Biol Cell, 22, 3645-3657. View more movies and download a PDF.

View the 'Motility of early endosomes' movie
(445Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 348Kb

Download the PC file - 555Kb

View the 'On the run loading and release of EE and dynein' movie

k. 'On the run' loading and release of EE and dynein
Loading: Anterograde EE (marked by mCherry-Rab5a, red) meets a retrograde dynein (labelled by GFP3-Dyn2, green). The dynein binds to the EE and changes the transport direction. Release: Dynein and EEs co-localise while moving in retrograde direction. An EE (red) moves in anterograde direction to the tip, which is mediated by kinesin-3. Time is given in seconds : milliseconds, the bars are given in micrometers. Reference: Schuster et al., 2011, PNAS, 108, 3618-3623. Download a PDF.

View the 'On the run loading and release of EE and dynein' movie
(97Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 115Kb

Download the PC file - 542Kb

View the 'Co-localisation of kinesin-3 and early endosomes' movie

l. Co-localisation of kinesin-3 and early endosomes
Kin3-GFP (green) was co-expressed with mCherry-Rab5a (red). The hypha was partially photo-bleached to visualise only the anterograde transport. Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Schuster et al., 2011, PNAS, 108, 3618-3623. Download a PDF.

View the 'Co-localisation of kinesin-3 and early endosomes' movie
(502Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 467Kb

Download the PC file -1.23Mb

4. The cytoskeleton in Ustilago maydis

View the 'Microtubule organisation in a hyphal cell' movie

a. Microtubule organisation in a hyphal cell
Microtubules form bundles that extend from the growing tip to the proximal septum, thereby providing continuous tracks that connect both cell poles. Note that individual microtubules can be very short and are not nucleated at the spindle pole body, a nuclear microtubule organising centre. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. The 3D reconstruction was built from a deconvolved Z-axis image stack. The bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Microtubule organisation in a hyphal cell' movie
(119Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 88Kb

Download the PC file - 396Kb

View the 'Microtubule dynamics in a hyphal cell' movie

b. Microtubule dynamics in a hyphal cell
Single microtubule undergoes frequent bending and waving whereas the bundles are less flexible. The movie was built from a deconvolved image series. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Microtubule dynamics in a hyphal cell' movie
(646Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 494Kb

Download the PC file - 1.35Mb

View the 'Microtubule organisation in a yeast-like cell' movie

c. Microtubule organisation in a yeast-like cell
Microtubules connect the growth region (top) with the mother cell (bottom). Microtubules have a tendency to form bundles and the microtubule array connects the cell poles. Like in hyphal cells, individual microtubules can be very short and are not nucleated at the spindle pole body, but are formed at cytoplasmic sites near the bud neck. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. The 3D reconstruction was built from a deconvolved Z-axis image stack. Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Microtubule organisation in a yeast-like cell' movie
(106Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 83Kb

Download the PC file - 318Kb

View the 'Microtubule dynamics in a yeast-like cell' movie

d. Microtubule dynamics in a yeast-like cell
Single microtubules polymerise and depolymerise. Dark patches on the microtubules (regions of less fluorescence called 'speckles'), can serve as 'landmarks' that demonstrate that (a) microtubules indeed elongate and (b) undergo short range to-and-fro motion. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. The movie was built from a deconvolved image series. The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Microtubule dynamics in a yeast-like cell' movie
(706Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 414Kb

Download the PC file - 787Kb

View the 'Microtubule organisation in a mitotic cell during late anaphase' movie

e. Microtubule organisation in a mitotic cell during late anaphase
The anaphase spindle extends from the daughter to the mother cell. Numerous astral microtubules are formed that position both spindle poles in the cell centres. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. The 3D reconstruction was built from a deconvolved Z-axis image stack. Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Microtubule organisation in a mitotic cell during late anaphase' movie
(106Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 83Kb

Download the PC file - 287Kb

View the 'Microtubule dynamics in a mitotic cell during anaphase' movie

f. Microtubule dynamics in a mitotic cell during anaphase
Long astral microtubules emanate from the spindle pole bodies and contact the cell cortex (cell periphery). Here cytoplasmic dynein pulls on the astral microtubules thereby elongating the spindle. The chromosomes (not visible) are located very close to the spindle pole bodies. Note that the behaviour of the spindle indicates counteracting forces exerted at both sides of the spindle. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. The time is given in seconds:milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Microtubule dynamics in a mitotic cell during anaphase' movie
(428Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 316Kb

Download the PC file - 654Kb

View the 'Dynamics of a microtubule aster during anaphase' movie

g. Dynamics of a microtubule aster during anaphase
The movie demonstrates the dynamic behaviour of the astral microtubules. Their interaction with dynein at the cellular cortex drives the motility of the spindle pole and thereby the segregation of the chromosomes. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. The movie was built from a deconvolved image series. The time is given in seconds:milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of a microtubule aster during anaphase' movie
(748Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 580Kb

Download the PC file - 771Kb

View the 'F-actin organisation in a hyphal U. maydis cell' movie

h. F-actin organisation in a hyphal U. maydis cell
F-actin is found as long peripheral filaments and brightly labelled peripheral patches. The cables provide 'tracks' for myosin motors, whereas the patches represent unordered F-actin accumulations that, due to their capacity to polymerise, are thought to propel endocytic vesicles from the cell periphery into the cytoplasm. Actin patches are concentrated at the hyphal tip indicating that this region is active in endocytosis. F-actinwas visualised by eGFP fused to a Lifeact-peptide. The 3D reconstruction was built from a deconvolved Z-axis image stack. The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'F-actin organisation in a hyphal U. maydis cell' movie
(173Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 116Kb

Download the PC file - 334Kb

View the 'Dynamics of F-actin patches at the tip of a hyphal cell' movie

i. Dynamics of F-actin patches at the tip of a hyphal cell
Actin patches are continuously formed at the cell periphery and are rapidly disappearing due to their disassembly and their motility out of the focal plane. F-actin was visualised by eGFP fused to a Lifeact-peptide. The movie was built from a deconvolved image series.The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of F-actin patches at the tip of a hyphal cell' movie
(375Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 371Kb

Download the PC file - 490Kb

View the 'Dynamics of F-actin in a yeast-like cell' movie

k. Dynamics of F-actin in a yeast-like cell
Actin patches are transiently visible. Note that the cables are stable peripheral structures. Patches occasionally move over some distances, indicating the motility of endocytic vesicles that are propelled by actin polymerisation. F-actin was visualised by eGFP fused to a Lifeact-peptide. The movie was built from a deconvolved image series. Time is given in seconds : milliseconds, the bars are given in micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of F-actin in a yeast-like cell' movie
(631Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 512Kb

Download the PC file - 928Kb

View the 'The cytoskeleton in yeast-like cells of Ustilago maydis' movie

l. The cytoskeleton in yeast-like cells of Ustilago maydis
F-actin was visualised by eGFP fused to a Lifeact-peptide. Microtubules were visualised by eGFP fused to the alpha-tubulin gene tub1. The movie was built from a deconvolved image series. The time is given in seconds:milliseconds; the bar represents micrometers. Reference: Schuster et al., 2011, EMBO J., 31, 214-327 Download a PDF.

View the 'The cytoskeleton in yeast-like cells of Ustilago maydis' movie
(215Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 199Kb

Download the PC file - 303Kb

5. Organization of the hyphal cell of Ustilago maydis

View the 'Dynamics of vacuoles in a hyphal cell of Ustilago maydis' movie

a. Dynamics of vacuoles in a hyphal cell of Ustilago maydis
The organelles remain almost stationary during the course of observation. Vacuoles are labelled by carboxypeptidase Y fused to a triple red fluorescent protein. Time is given in seconds:milliseconds. The bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of vacuoles in a hyphal cell of Ustilago maydis' movie
(511Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 369Kb

Download the PC file - 959Kb

View the 'Vacuole distribution in a yeast-like cell of Ustilago maydis' movie

b. Vacuole distribution in a yeast-like cell of Ustilago maydis
The organelles are spherical and evenly scattered throughout the cell. Note that the number and size of the vacuoles can vary and depends on unknown environmental conditions. Vacuoles were labelled by the dye Cell Tracker Blue (Molecular Probes). The 3D reconstruction was built from a deconvolved Z-axis image stack Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Vacuole distribution in a yeast-like cell of Ustilago maydis' movie
(101Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 116Kb

Download the PC file - 591Kb

View the 'Dynamics of the endoplasmic reticulum' movie

c. Dynamics of the endoplasmic reticulum in a hyphal cell of Ustilago maydis
The network remains mainly stationary. Occasionally long-range motility occurs (one event indicated by arrowhead). The endoplasmic reticulum and the nuclear envelope was visualised by expressing eGFP, N-terminally fused to a calreticulin signal peptide and C-terminally fused to the retention signal HDEL.Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of the endoplasmic reticulum' movie
(1.04Mb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 581Kb

Download the PC file - 1.49Mb

View the 'Dynamics of the endoplasmic reticulum' movie

d. Dynamics of the endoplasmic reticulum in a yeast-like cell of Ustilago maydis
The peripheral network is undergoing thermal flickering. In addition, directed motility of tubules occurs that is mediated by microtubule motors. The endoplasmic reticulum was visualised by expressing eGFP, N-terminally fused to a calreticulin signal peptide and C-terminally fused to the retention signal HDEL. Time is given in seconds : milliseconds, the bar is given in micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of the endoplasmic reticulum' movie
(486Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 447Kb

Download the PC file - 1003Kb

View the 'Motility of the endoplasmic reticulum' movie

e. Motility of the endoplasmic reticulum and microtubules in Ustilago maydis
Bi-directional motility occurs around the microtubule tracks. The endoplasmic reticulum and the nuclear envelope was visualised by expressing eGFP, N-terminally fused to a calreticulin signal peptide and C-terminally fuse to the retention signal HDEL. The microtubules were visualised by monomeric Cherry fused to the alpha-tubulin gene tub1. The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Motility of the endoplasmic reticulum' movie
(455Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 802Kb

Download the PC file - 1.07Mb

View the 'Dynamics of peroxisomes in a hyphal cell of Ustilago maydis' movie

f. Dynamics of peroxisomes in a hyphal cell of Ustilago maydis
Most peroxisomes are stationary or undergo Brownian motion. Occasionally, peroxisomes show rapid bi-directional motility. Note that the halos result from peroxisomes that are not in focus. Peroxisomes were labelled by eGFP fused to the import tri-peptide SKL. The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of peroxisomes in a hyphal cell of Ustilago maydis' movie
(250Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 289Kb

Download the PC file - 584Kb

View the 'Dynamics of mitochondria in a hyphal cell of Ustilago maydis' movie

g. Dynamics of mitochondria in a hyphal cell of Ustilago maydis
Most organelles are stationary or undergo Brownian motion. Occasionally, mitochondria show short range motility (arrowhead). Mitochondria were labelled by the U. maydis-specific matrix protein Lga2 fused to eGFP. The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Dynamics of mitochondria in a hyphal cell of Ustilago maydis' movie
(384Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 445Kb

Download the PC file - 1006Kb

View the 'The nucleus in a yeast-like cell of Ustilago maydis' movie

h. The nucleus in a yeast-like cell of Ustilago maydis
The nucleus was labelled by a triple RFP tag fused to a nuclear localisation signal. The 3D reconstruction was built from a deconvolved Z-axis image stack. Bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'The nucleus in a yeast-like cell of Ustilago maydis' movie
(88Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 98Kb

Download the PC file - 790Kb

View the 'Recovery of fluorescence in the plasma membrane' movie

i. Recovery of fluorescence in the plasma membrane after photo-bleaching an Ustilago maydis cell
The fluorescence was bleached by a 405 nm laser pulse (red circle, bleach).With time the un-bleached molecules from neighbouring regions of the plasma membrane diffuse into the darkened area. This indicates that molecules can “swim” within the bi-layer, which illustrates the “fluid mosaic model” of bio-membranes. The plasma membrane was labelled by a fusion protein of eGFP and a Sso1-like syntaxin. The time is given in seconds: milliseconds; the bar represents micrometers. Reference: Steinberg and Schuster, 2011, Fungal Biol. Rev., 25, 14-37. Download a PDF.

View the 'Recovery of fluorescence in the plasma membrane' movie
(1001Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.67Mb

Download the PC file - 1.96Mb

6. Additional Movies

View the 'Infection of maize leave tissue by Ustilago maydis' movie

a. Infection of maize leave tissue by Ustilago maydis (3 days post infection)
The fungus growth intracellular. Fungal hyphae were stained with agglutinin-AF488 (green), the plant cell walls were visualized by propidium iodide (red). The bar represents micrometers. Reference: Treitschke et al., 2010, Plant Cell, 22, 2476-2494. More movies and download a PDF.

View the 'Infection of maize leave tissue by Ustilago maydis' movie
(682Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.04Mb

Download the PC file - 1.21Mb

View the 'Infection of maize leave tissue' movie

b. Infection of maize leave tissue by a Ustilago maydis mutant that was deleted in a class V chitin synthase (3 days post infection)
In the absence of the chitin synthase the fungus swells and cannot invade the plant. Fungal hyphae were stained with agglutinin-AF488 (green), the plant cell walls were visualized by propidium iodide (red). The bar represents micrometers. Reference: Treitschke et al., 2010, Plant Cell, 22, 2476-2494. More movies and download a PDF.

View the 'Infection of maize leave tissue' movie
(193Kb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 188Kb

Download the PC file - 365Kb

View the 'Crawling pseudopdia of wall-less cell fragments' movie

c. Crawling pseudopdia of wall-less cell fragments of a Neurospora crassa mutant
The mutant (called SLIME) is defective in β-D-glucan synthase (fz;sg;os-1; FGSC 1118). Cells were attached to a protamine-sulfate coated cover slip. This movie demonstrates that the cell has the capacity to expand without the cell-wall. The movie runs at real time, the elapsed time is given in minutes:seconds. Reference: Steinberg 2007, Euk. Cell, 6, 351-360. Download a PDF.

View the 'Crawling pseudopdia of wall-less cell fragments' movie
(1.49Mb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 893Kb

Download the PC file - 1.14Mb

View the 'Waving motility of the wall-less Neurospora crassa SLIME mutant' movie

d. Waving motility of the wall-less Neurospora crassa SLIME mutant
Motility of a wall-less N. crassa mutant, defective in β-D-glucan synthase (fz;sg;os-1; FGSC 1118) occurs in phosphate buffered saline. This movie demonstrates that the cell can exert turgor0independent motility that is reminiscent of amoeboid motion in animal and protozoa cells. The movie runs at real time, the elapsed time is given in minutes:seconds. Reference: Steinberg 2007, Euk. Cell, 6, 351-360. Download a PDF.

View the 'Waving motility of the wall-less Neurospora crassa SLIME mutant' movie
(1.79Mb – Opens in a new window)

To download please right click and save link as.

Download the Mac file - 1.65Mb

Download the PC file - 2.00Mb