Scm3, an essential Saccharomyces cerevisiae centromere protein required for G2/M progression and Cse4 localization.
A universal mark of centromeric chromatin is its packaging by a variant of histone H3 known as centromeric H3 (CenH3). The mechanism by which CenH3s are incorporated specifically into centromere DNA or the specialized function they serve there is not known. In a genetic approach to identify factors involved in CenH3 deposition, we screened for dosage suppressors of a temperature-sensitive cse4 allele in Saccharomyces cerevisiae (Cse4 is the S. cerevisiae CenH3). Independent screens yielded ORF YDL139C, which we named SCM3. Dosage suppression by SCM3 was specific for alleles affecting the histone fold domain of Cse4. Copurification and two-hybrid studies showed that Scm3 and Cse4 interact in vivo, and chromatin immunoprecipitation revealed that Scm3, like Cse4, is found associated with centromere DNA. Scm3 contains two essential protein domains, a Leu-rich nuclear export signal and a heptad repeat domain that is widely conserved in fungi. A conditional scm3 allele was generated to allow us to deplete Scm3. Upon Scm3 depletion, cells undergo a Mad2-dependent G2/M arrest, and centromere localization of Cse4 is perturbed. We suggest that S. cerevisiae Scm3 defines a previously undescribed family of fungal kinetochore proteins important for CenH3 localization.
Stoler
Sam
S
Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA 01003, USA.
Rogers
Kelly
K
Weitze
Scott
S
Morey
Lisa
L
Fitzgerald-Hayes
Molly
M
Baker
Richard E
RE
D000483:Alleles; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001499:Bayes Theorem; D002503:Centromere / Q000737:chemistry / Q000235:genetics*; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D016195:G2 Phase; D005816:Genetic Complementation Test; D006388:Hemagglutinins / Q000737:chemistry; D007930:Leucine / Q000737:chemistry; D008938:Mitosis; D008957:Models, Genetic; D008969:Molecular Sequence Data; D049790:Nuclear Export Signals; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D013912:Threonine / Q000378:metabolism; D020798:Two-Hybrid System Techniques
C092746:CSE4 protein, S cerevisiae; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006388:Hemagglutinins; D049790:Nuclear Export Signals; D029701:Saccharomyces cerevisiae Proteins; C521684:Scm3 protein, S cerevisiae; D013912:Threonine; D007930:Leucine