SBA1 encodes a yeast hsp90 cochaperone that is homologous to vertebrate p23 proteins.
The Saccharomyces cerevisiae SBA1 gene was cloned by PCR amplification from yeast genomic DNA following its identification as encoding an ortholog of human p23, an Hsp90 cochaperone. The SBA1 gene product is constitutively expressed and nonessential, although a disruption mutant grew more slowly than the wild type at both 18 and 37 degreesC. A double deletion of SBA1 and STI1, encoding an Hsp90 cochaperone, displayed synthetic growth defects. Affinity isolation of histidine-tagged Sba1p (Sba1(His6)) after expression in yeast led to coisolation of Hsp90 and the cyclophilin homolog Cpr6. Using an in vitro assembly assay, purified Sba1(His6) bound to Hsp90 only in the presence of adenosine 5'-O-(3-thiotriphosphate) or adenyl-imidodiphosphate. Furthermore, interaction between purified Sba1(His6) and Hsp90 in yeast extracts was inhibited by the benzoquinoid ansamycins geldanamycin and macbecin. The in vitro assay was also used to identify residues in Hsp90 that are important for complex formation with Sba1(His6), and residues in both the N-terminal nucleotide binding domain and C-terminal half were characterized. In vivo analysis of known Hsp90 substrate proteins revealed that Sba1 loss of function had only a mild effect on the activity of the tyrosine kinase v-Src and steroid hormone receptors.
Fang
Y
Y
Department of Cell Biology and Anatomy, Mount Sinai Medical Center, New York, New York 10029, USA.
Fliss
A E
AE
Rao
J
J
Caplan
A J
AJ
D000595:Amino Acid Sequence; D000818:Animals; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D015688:Oncogene Protein pp60(v-src) / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics*; D000071876:Prostaglandin-E Synthases; D011944:Receptors, Androgen / Q000378:metabolism; D011980:Receptors, Progesterone / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins
D005656:Fungal Proteins; D018841:HSP90 Heat-Shock Proteins; D018832:Molecular Chaperones; D010750:Phosphoproteins; D011944:Receptors, Androgen; D011980:Receptors, Progesterone; C113025:SBA1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015688:Oncogene Protein pp60(v-src); C490849:PTGES3 protein, human; D000071876:Prostaglandin-E Synthases