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Mol Cell Biol, July 1998, p. 3727-3734, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
SBA1 Encodes a Yeast Hsp90 Cochaperone
That Is Homologous to Vertebrate p23 Proteins
Yifang
Fang,
Albert E.
Fliss,
Jie
Rao, and
Avrom J.
Caplan*
Department of Cell Biology and Anatomy, Mount
Sinai Medical Center, New York, New York 10029
Received 26 November 1997/Returned for modification 20 January
1998/Accepted 31 March 1998
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°C. A double deletion of SBA1
and STI1, encoding an Hsp90 cochaperone, displayed
synthetic growth defects. Affinity isolation of histidine-tagged Sba1p
(Sba1His6) after expression in yeast led to coisolation of
Hsp90 and the cyclophilin homolog Cpr6. Using an in vitro assembly
assay, purified Sba1His6 bound to Hsp90 only in the
presence of adenosine 5'-O-(3-thiotriphosphate) or
adenyl-imidodiphosphate. Furthermore, interaction between purified Sba1His6 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 Sba1His6, 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.
*
Corresponding author. Mailing address: Department of
Cell Biology and Anatomy, Mount Sinai Medical Center, One Gustave L. Levy Place, New York, NY 10029. Phone: (212) 241-6563. Fax: (212) 860-1174. E-mail: caplan{at}msvax.mssm.edu.
Mol Cell Biol, July 1998, p. 3727-3734, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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