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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

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

hyphal extension (Natalia Requena)

Wild type

DkinA

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


Myosin

Actin

MT

Polarised growth and the Spitzenkörper

Conventional Kinesin

B. Robertson,

Athens GA


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


Polarity mutants in S. pombe

Dtea1 and Dtea2

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


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


KipA deletion affects polarized growth

(Sven Konzack)

Wild type

kipA (tea2) mutant


wild type

kipA-deletion

KipA deletion affects Spitzenkörper position


wild type

DkipA

KipA is required for microtuble organization

in the tip

Difference to S. pombeDtea2


GFP

GFP

KipA localization

Sven Konzack


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


KipA

KipA

KipA

KipA

Microtubule plus end

TeaA?

Hestermann et al., 2002


KipA and polarized growth

DkipA

Wild type

KipA

TeaA?

Conventional Kinesin

Putative TeaA receptor


wild type

DkipA

DteaA

DkipA and DteaA display distinct phenotypes


AlpA?

KipA

KipA

KipA

KipA

Microtubule plus end

Does KipA transport other cargoes?

Hestermann et al., 2002


GFP-MT

mRFP1-AlpA

0

2:27

Localization of AlpA in A. nidulans(Cathrin Enke)

Mitosis

GFP-AlpA


WT DalpA

Curved hyphae

25°C

37°C

42°C

Deletion of alpA


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


ApsA

TeaA?

AlpA

NudF

NudA

KipA

KinA

Different cortical interactions for

nuclear migration and polarized growth


mRFP-Microtubules

GFP-KipA

Microtubule polarity in A. nidulans

Microtubule

tracks are used

several times


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


Nuclei

Mitos

Nuclei

Mitos

BFP

sGFP

mRFP1

Tools

Fluorescent proteins

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


High-throughput tagging (Gateway)

Gene of

interest

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


Anne Blumenstein

Kay Vienken

Alex Schütz

Sven Konzack

Cathrin Enke

Daniel Veith

Nicole Scherr

Patricia Rischitor

[email protected]


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