Sfi1p has conserved centrin-binding sites and an essential function in budding yeast spindle pole body duplication

doi:10.1083/jcb.200307064

Published online 22 September 2003. doi:10.1083/jcb.200307064

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© The Rockefeller University Press, 0021-9525/2003/9/1211 $5.00
The Journal of Cell Biology, Volume 162, Number 7, 1211-1221

Article

Sfi1p has conserved centrin-binding sites and an essential function in budding yeast spindle pole body duplication

John V. Kilmartin

Medical Research Council (MRC) Laboratory of Molecular Biology, Cambridge CB2 2QH, UK

Address correspondence to J.V. Kilmartin, MRC Laboratory of Molecular Biology, Hills Rd., Cambridge CB2 2QH, UK. Tel.: 44-1223-402242. Fax: 44-1223-412142. email: jvk{at}mrc-lmb.cam.ac.uk

Centrins are calmodulin-like proteins present in microtubule-organizingcenters. The Saccharomyces cerevisiae centrin, Cdc31p, was functionallytagged with a single Z domain of protein A, and used in pull-downexperiments to isolate Cdc31p-binding proteins. One of these,Sfi1p, localizes to the half-bridge of the spindle pole body(SPB), where Cdc31p is also localized. Temperature-sensitivemutants in SFI1 show a defect in SPB duplication and geneticinteractions with cdc31-1. Sfi1p contains multiple internalrepeats that are also present in a Schizosaccharomyces pombeprotein, which also localizes to the SPB, and in several humanproteins, one of which localizes close to the centriole region.Cdc31p binds directly to individual Sfi1 repeats in a 1:1 ratio,so a single molecule of Sfi1p binds multiple molecules of Cdc31p.The centrosomal human protein containing Sfi1 repeats also bindscentrin in the repeat region, showing that this centrin-bindingmotif is conserved.

Key Words: SPB; duplication; Sfi1p; Cdc31p; centrin

Abbreviations used in this paper: prA, protein A; SPB, spindlepole body; ZCdc31p, Cdc31p containing a single Z domain of proteinA at the NH₂ terminus.

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