Alaamery Manal A, Hoffman Charles S
Biology Department, Boston College, Chestnut Hill, Massachusetts 02467, USA.
Genetics. 2008 Apr;178(4):1927-36. doi: 10.1534/genetics.107.086165.
The fission yeast Schizosaccharomyces pombe senses environmental glucose through a cAMP-signaling pathway. Elevated cAMP levels activate protein kinase A (PKA) to inhibit transcription of genes involved in sexual development and gluconeogenesis, including the fbp1(+) gene, which encodes fructose-1,6-bisphosphatase. Glucose-mediated activation of PKA requires the function of nine glucose-insensitive transcription (git) genes, encoding adenylate cyclase, the PKA catalytic subunit, and seven "upstream" proteins required for glucose-triggered adenylate cyclase activation. We describe the cloning and characterization of the git10(+) gene, which is identical to swo1(+) and encodes the S. pombe Hsp90 chaperone protein. Glucose repression of fbp1(+) transcription is impaired by both git10(-) and swo1(-) mutant alleles of the hsp90(+) gene, as well as by chemical inhibition of Hsp90 activity and temperature stress to wild-type cells. Unlike the swo1(-) mutant alleles, the git10-201 allele supports cell growth at 37 degrees , while severely reducing glucose repression of an fbp1-lacZ reporter, suggesting a separation-of-function defect. Sequence analyses of three swo1(-) alleles and the one git10(-) allele indicate that swo1(-) mutations alter core functional domains of Hsp90, while the git10(-) mutation affects the Hsp90 central domain involved in client protein binding. These results suggest that Hsp90 plays a specific role in the S. pombe glucose/cAMP pathway.
裂殖酵母粟酒裂殖酵母通过cAMP信号通路感知环境中的葡萄糖。cAMP水平升高会激活蛋白激酶A(PKA),从而抑制参与有性发育和糖异生的基因转录,包括编码果糖-1,6-二磷酸酶的fbp1(+)基因。葡萄糖介导的PKA激活需要九个葡萄糖不敏感转录(git)基因的功能,这些基因编码腺苷酸环化酶、PKA催化亚基以及葡萄糖触发的腺苷酸环化酶激活所需的七种“上游”蛋白。我们描述了git10(+)基因的克隆和特征,该基因与swo1(+)相同,编码粟酒裂殖酵母Hsp90伴侣蛋白。hsp90(+)基因的git10(-)和swo1(-)突变等位基因,以及对野生型细胞的Hsp90活性的化学抑制和温度胁迫,都会损害fbp1(+)转录的葡萄糖抑制作用。与swo1(-)突变等位基因不同,git10-201等位基因在37摄氏度时支持细胞生长,同时严重降低fbp1-lacZ报告基因的葡萄糖抑制作用,表明存在功能分离缺陷。对三个swo1(-)等位基因和一个git10(-)等位基因的序列分析表明,swo1(-)突变改变了Hsp90的核心功能域,而git10(-)突变影响了参与客户蛋白结合的Hsp90中央结构域。这些结果表明,Hsp90在粟酒裂殖酵母葡萄糖/cAMP途径中发挥特定作用。