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Microtubules and polarized growth in Aspergillus nidulans. Reinhard Fischer Max-Planck-Institute for Terrestrial Microbiology. Myosin. Actin. MT. Polarised growth and the Spitzenkörper. B. Robertson, Athens GA. Conventional kinesin is required for

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Microtubules and polarized growth in Aspergillus nidulans

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Microtubules and polarized growth in aspergillus nidulans

Microtubules and polarized growth in Aspergillus nidulans

Reinhard Fischer

Max-Planck-Institute for Terrestrial Microbiology


Microtubules and polarized growth in aspergillus nidulans

Myosin

Actin

MT

Polarised growth and the Spitzenkörper

B. Robertson,

Athens GA


Microtubules and polarized growth in aspergillus nidulans

Conventional kinesin is required for

hyphal extension (Natalia Requena)

Wild type

DkinA

Requena et al., 2001, Mol. Mirobiol. 42: 121-132


Microtubules and polarized growth in aspergillus nidulans

Myosin

Actin

MT

Polarised growth and the Spitzenkörper

Conventional Kinesin

B. Robertson,

Athens GA


Microtubules and polarized growth in aspergillus nidulans

Maekawa et al., 2003, EMBO J. 22: 438-449

Cortical interactions of microtubules:

Nuclear migration in S. cerevisiae

Num1 (ApsA)

Yamamoto et al., 2003, JCS 116:4501-4512


Microtubules and polarized growth in aspergillus nidulans

Polarity mutants in S. pombe

Dtea1 and Dtea2

Browning et al., 2000, JCB, 151:15-27


Microtubules and polarized growth in aspergillus nidulans

Microtubule organization inDtea1 and Dtea2

strains

WT

Dtea1

Dtea2

Browning et al., 2000, JCB, 151:15-27

Mata and Nurse, 1997, Cell 89:939-949

Tea1 destabilizes microtubules

Tea2 stabilizes microtubules


Microtubules and polarized growth in aspergillus nidulans

KipA deletion affects polarized growth

(Sven Konzack)

Wild type

kipA (tea2) mutant


Microtubules and polarized growth in aspergillus nidulans

wild type

kipA-deletion

KipA deletion affects Spitzenkörper position


Microtubules and polarized growth in aspergillus nidulans

wild type

DkipA

KipA is required for microtuble organization

in the tip

Difference to S. pombeDtea2


Microtubules and polarized growth in aspergillus nidulans

GFP

GFP

KipA localization

Sven Konzack


Microtubules and polarized growth in aspergillus nidulans

mRFP1-Microtubules

GFP-KipA

Microtubule dynamics

Xin Xiang, Ron Morris

KipA is delivered

in the tip

Morris, Curr. Opin. Cell Biol. 15:54-59, 2003

Efimov, Mol. Biol. Cell 14:871-888, 2003

Zhang et al., Mol. Biol. Cell 15: , 2003


Microtubules and polarized growth in aspergillus nidulans

KipA

KipA

KipA

KipA

Microtubule plus end

TeaA?

Hestermann et al., 2002


Microtubules and polarized growth in aspergillus nidulans

KipA and polarized growth

DkipA

Wild type

KipA

TeaA?

Conventional Kinesin

Putative TeaA receptor


Microtubules and polarized growth in aspergillus nidulans

wild type

DkipA

DteaA

DkipA and DteaA display distinct phenotypes


Microtubules and polarized growth in aspergillus nidulans

AlpA?

KipA

KipA

KipA

KipA

Microtubule plus end

Does KipA transport other cargoes?

Hestermann et al., 2002


Localization of alpa in a nidulans cathrin enke

GFP-MT

mRFP1-AlpA

0

2:27

Localization of AlpA in A. nidulans(Cathrin Enke)

Mitosis

GFP-AlpA


Microtubules and polarized growth in aspergillus nidulans

WT DalpA

Curved hyphae

25°C

37°C

42°C

Deletion of alpA


Microtubules and polarized growth in aspergillus nidulans

Microtubule plus end

Curved hyphae

KipA

AlpA (XMAP)

NudF (LIS1)

Cortical protein: TeaA

No curved hyphae

NudA (dynein)

Cortical protein: ApsA

Hestermann et al., 2002


Microtubules and polarized growth in aspergillus nidulans

ApsA

TeaA?

AlpA

NudF

NudA

KipA

KinA

Different cortical interactions for

nuclear migration and polarized growth


Microtubules and polarized growth in aspergillus nidulans

mRFP-Microtubules

GFP-KipA

Microtubule polarity in A. nidulans

Microtubule

tracks are used

several times


Microtubules and polarized growth in aspergillus nidulans

Kinesins

in A. nidulans

  • BimCMitosis (R. Morris)

  • KlpAMitosis (R. Morris)

  • KipBMicrotubule dynamics,

  • Mitosis (P. Rischitor)

  • KinA Tip extension (N. Requena)

  • KipA Directionality (S. Konzack)

  • Two Unc104 homologues plus four

  • others to be studied

Interphase

Mitosis

Interphase after septation

103

137

(240/50)

56

123

285

117

7

56

(112/16)

(240/29)

(292/42)

Microtubule organization

in A. nidulans


Microtubules and polarized growth in aspergillus nidulans

Nuclei

Mitos

Nuclei

Mitos

BFP

sGFP

mRFP1

Tools

Fluorescent proteins

Toews et al., 2004, Curr. Genet. online


Microtubules and polarized growth in aspergillus nidulans

High-throughput tagging (Gateway)

Gene of

interest

Toews et al., 2004, Curr. Genet. online


Microtubules and polarized growth in aspergillus nidulans

Anne Blumenstein

Kay Vienken

Alex Schütz

Sven Konzack

Cathrin Enke

Daniel Veith

Nicole Scherr

Patricia Rischitor

[email protected]


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