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Published online 22 January 2001. doi:10.1083/jcb.152.2.251
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© The Rockefeller University Press, 0021-9525/2001//251 $5.00
The Journal of Cell Biology, Volume 152, Number 2, , 2001 251-262

Original Article

Cortical Num1p Interacts with the Dynein Intermediate Chain Pac11p and Cytoplasmic Microtubules in Budding Yeast



Marian Farkasovskya and Hans Küntzela

a Max-Planck Institute for Experimental Medicine, D-37075 Göttingen, Germany
Max-Planck-Institut für Experimentelle Medizin, Hermann-Rein-Strasse 3, D-37075 Göttingen, Germany.49-0551-3899-35249-0551-3899-278

kuentzel{at}em.mpg.de

Num1p, a cortical 313-kD protein, controls cytoplasmic microtubule (cMT) functions and nuclear migration through the bud neck in anaphase cells. A green fluorescent protein (GFP)-Num1p fusion protein localizes at the bud tip and the distal mother pole of living cells, apparently forming cMT capture sites at late anaphase. In addition, galactose-induced GFP-Num1p is seen at the bud neck and in lateral regions of the mother cortex. The bud tip location of Num1p depends on Bni1p but does not require Kar9p, Dyn1p, or cMTs, whereas cMT contacts with polar Num1p dots are reduced in cells lacking Dyn1p. Num1p associates with the dynein intermediate chain Pac11p in the presence of Dyn1p, and with the {alpha}-tubulin Tub3p, as shown by coimmune precipitation of tagged proteins. Num1p also forms a complex with Bni1p and Kar9p, although Num1p is not required for Bni1p- and Kar9p-dependent nuclear migration to the bud neck in preanaphase cells.

Our data suggest that Num1p controls nuclear migration during late anaphase by forming dynein-interacting cortical cMT capture sites at both cellular poles. In addition, Num1p may transiently cooperate with an associated Bni1p–Kar9p complex at the bud tip of early anaphase cells.

Key Words: microtubule • cytoskeleton • nuclear migration • dynein • yeast

© 2001 The Rockefeller University Press

Marian Farkasovsky's present address is Slovak Academy of Sciences, Institute of Molecular Biology, 84251 Bratislava, Slovakia.

Abbreviations used in this paper: cMT, cytoplasmic microtubule; GFP, green fluorescent protein; HA, hemagglutinin; Myc, c-Myc; PH, pleckstrin homology; SPB, spindle pole body; yEGFP, yeast-enhanced GFP; YPD, yeast extract/peptone plus dextrose.


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