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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

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slide1

Microtubules and polarized growth in Aspergillus nidulans

Reinhard Fischer

Max-Planck-Institute for Terrestrial Microbiology

slide2

Myosin

Actin

MT

Polarised growth and the Spitzenkörper

B. Robertson,

Athens GA

slide3

Conventional kinesin is required for

hyphal extension (Natalia Requena)

Wild type

DkinA

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

slide4

Myosin

Actin

MT

Polarised growth and the Spitzenkörper

Conventional Kinesin

B. Robertson,

Athens GA

slide5

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

slide6

Polarity mutants in S. pombe

Dtea1 and Dtea2

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

slide7

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

slide8

KipA deletion affects polarized growth

(Sven Konzack)

Wild type

kipA (tea2) mutant

slide9

wild type

kipA-deletion

KipA deletion affects Spitzenkörper position

slide10

wild type

DkipA

KipA is required for microtuble organization

in the tip

Difference to S. pombeDtea2

slide11

GFP

GFP

KipA localization

Sven Konzack

slide12

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

slide13

KipA

KipA

KipA

KipA

Microtubule plus end

TeaA?

Hestermann et al., 2002

slide14

KipA and polarized growth

DkipA

Wild type

KipA

TeaA?

Conventional Kinesin

Putative TeaA receptor

slide15

wild type

DkipA

DteaA

DkipA and DteaA display distinct phenotypes

slide16

AlpA?

KipA

KipA

KipA

KipA

Microtubule plus end

Does KipA transport other cargoes?

Hestermann et al., 2002

slide18

WT DalpA

Curved hyphae

25°C

37°C

42°C

Deletion of alpA

slide19

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

slide20

ApsA

TeaA?

AlpA

NudF

NudA

KipA

KinA

Different cortical interactions for

nuclear migration and polarized growth

slide21

mRFP-Microtubules

GFP-KipA

Microtubule polarity in A. nidulans

Microtubule

tracks are used

several times

slide22

Kinesins

in A. nidulans

  • BimC Mitosis (R. Morris)
  • KlpA Mitosis (R. Morris)
  • KipB Microtubule 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

slide23

Nuclei

Mitos

Nuclei

Mitos

BFP

sGFP

mRFP1

Tools

Fluorescent proteins

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

slide24

High-throughput tagging (Gateway)

Gene of

interest

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

slide25

Anne Blumenstein

Kay Vienken

Alex Schütz

Sven Konzack

Cathrin Enke

Daniel Veith

Nicole Scherr

Patricia Rischitor

[email protected]

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