Wickner R B
Section on Genetics of Simple Eukaryotes, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1988 Aug;85(16):6007-11. doi: 10.1073/pnas.85.16.6007.
The MAK16 gene was first defined as a gene whose mutation resulted in loss of M1 double-stranded RNA virus-like particles. The mak16-1 mutation also produces temperature-sensitive cell growth. We report here that mak16-1 cells arrest at the nonpermissive temperature in G1 phase, such that they are mating competent. We sequenced the MAK16 gene and found an open reading frame of 306 amino acids encoding a predicted protein of Mr 35,694. Two typical nuclear localization signal sequences were found. MAK16-LacZ fusion proteins that include one of these putative signals entered the nucleus, while unfused beta-galactosidase did not, as judged by subcellular fractionation experiments. In the C-terminal third of the MAK16 open reading frame is an acidic region in which 25 of 41 residues are either glutamate or aspartate. This region contains potential phosphorylation sites for "casein kinases," protein kinases specific for serine or threonine residues in an acidic environment.
MAK16基因最初被定义为一个其突变导致M1双链RNA病毒样颗粒缺失的基因。mak16 - 1突变也会产生温度敏感型细胞生长。我们在此报告，mak16 - 1细胞在非允许温度下停滞于G1期，因此它们具有交配能力。我们对MAK16基因进行了测序，发现一个306个氨基酸的开放阅读框，编码一个预测分子量为35,694的蛋白质。发现了两个典型的核定位信号序列。通过亚细胞分级实验判断，包含其中一个推定信号的MAK16 - LacZ融合蛋白进入了细胞核，而未融合的β - 半乳糖苷酶则没有。在MAK16开放阅读框的C端三分之一区域是一个酸性区域，其中41个残基中有25个是谷氨酸或天冬氨酸。该区域包含“酪蛋白激酶”的潜在磷酸化位点，酪蛋白激酶是在酸性环境中对丝氨酸或苏氨酸残基具有特异性的蛋白激酶。
