Watanabe Y, Yamamoto M
Department of Biophysics and Biochemistry, School of Science, University of Tokyo, Japan.
Mol Cell Biol. 1996 Feb;16(2):704-11. doi: 10.1128/MCB.16.2.704.
The Schizosaccharomyces pombe pcr1 gene encodes a bZIP protein that apparently belongs to the cyclic AMP response element (CRE)-binding protein/activating transcription factor family. The deduced pcr1 gene product consists of 171 amino acid residues and is most similar to the mammalian CRE-BP1. A glutathione S-transferase-Pcr1 fusion protein produced in Escherichia coli was able to bind specifically to the CRE motif in vitro. Analysis with anti-Pcr1 serum suggested that Pcr1 is included in the major CRE-binding factors present in the S. pombe cell extract. Disruption of the pcr1 gene was not lethal, but the disruptant showed cold-sensitive growth on rich medium. The disruptant was also inefficient in mating and sporulation, though it was not completely sterile. Expression of the ste11 gene, which encodes a key transcription factor for sexual development, was greatly reduced in the disruptant, and overexpression of ste11+ suppressed the deficiency of the pcr1 disruptant in sexual development. It has been shown that expression of ste11 is negatively regulated by cyclic AMP-dependent protein kinase (PKA) and that the loss of PKA activity results in ectopic sexual development. Disruption of pcr1 blocked ectopic sexual development. Furthermore, disruption of pcr1 reduced expression of fbp1, a glucose-repressible gene negatively regulated by PKA. These results suggest that Pcr1 is a putative transcriptional regulator whose activity may be controlled by PKA. Alternatively, its activity may be independent of PKA, and full induction of ste11 and fbp1 expression requires the function of Pcr1 in addition to elimination of the repression by PKA.
粟酒裂殖酵母的pcr1基因编码一种bZIP蛋白,该蛋白显然属于环磷酸腺苷反应元件(CRE)结合蛋白/激活转录因子家族。推导的pcr1基因产物由171个氨基酸残基组成,与哺乳动物的CRE-BP1最为相似。在大肠杆菌中产生的谷胱甘肽S-转移酶-Pcr1融合蛋白能够在体外特异性结合CRE基序。用抗Pcr1血清进行的分析表明,Pcr1包含在粟酒裂殖酵母细胞提取物中存在的主要CRE结合因子中。pcr1基因的破坏并非致命,但破坏株在丰富培养基上表现出冷敏感生长。该破坏株在交配和孢子形成方面也效率低下,尽管它并非完全不育。编码有性发育关键转录因子的ste11基因在破坏株中的表达大大降低,而ste11+的过表达抑制了pcr1破坏株在有性发育方面的缺陷。已经表明,ste11的表达受环磷酸腺苷依赖性蛋白激酶(PKA)负调控,PKA活性的丧失导致异位有性发育。pcr1的破坏阻止了异位有性发育。此外,pcr1的破坏降低了fbp1的表达,fbp1是一个受PKA负调控的葡萄糖可抑制基因。这些结果表明,Pcr1是一种假定的转录调节因子,其活性可能受PKA控制。或者,其活性可能独立于PKA,并且ste11和fbp1表达的完全诱导除了消除PKA的抑制作用外还需要Pcr1的功能。