Myre Michael A, O'Day Danton H
Department of Biology, University of Toronto at Mississauga, 3359 Mississauga Rd., Rm. 3030, Mississauga, ON, Canada L5L 1C6.
Biochim Biophys Acta. 2004 Nov 18;1675(1-3):192-7. doi: 10.1016/j.bbagen.2004.08.002.
A search of the Dictyostelium genome project database (http://dictybase.org/db/cgi-bin/blast.pl) with nucleomorphin, a protein that regulates the nuclear number, predicted it to be encoded by a larger gene containing a putative breast cancer carboxy-terminus domain (BRCT). Using RT-PCR, Northern and Western blotting we have identified a differentially expressed, 2318 bp cDNA encoding a protein isoform of Dictyostelium NumA with an apparent molecular weight of 70 kDa that we have called NumB. It contains a single amino-terminal BRCT-domain spanning residues 125-201. Starvation of shaking cultures reduces NumA expression by approximately 88+/-5.6%, whereas NumB expression increases approximately 35+/-3.5% from vegetative levels. NumC, a third isoform that is also expressed during development but not growth, remains to be characterized. These findings suggest NumB may be a member of the BRCT-domain containing cell cycle checkpoint proteins.
利用调节核数量的蛋白质核仁素对盘基网柄菌基因组计划数据库(http://dictybase.org/db/cgi-bin/blast.pl)进行搜索,预测它由一个更大的基因编码,该基因含有一个假定的乳腺癌羧基末端结构域(BRCT)。通过逆转录聚合酶链反应(RT-PCR)、Northern印迹法和Western印迹法,我们鉴定出一个差异表达的2318 bp cDNA,其编码盘基网柄菌NumA的一种蛋白质异构体,表观分子量为70 kDa,我们将其命名为NumB。它包含一个单一的氨基末端BRCT结构域,跨越第125 - 201位残基。振荡培养物饥饿处理使NumA表达降低约88±5.6%,而NumB表达从营养生长水平增加约35±3.5%。NumC是第三种异构体,在发育过程中表达但在生长过程中不表达,其特征还有待确定。这些发现表明NumB可能是含有BRCT结构域的细胞周期检查点蛋白家族的一员。