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酿酒酵母的ACP2基因编码一种必需的类HMG1蛋白。

The Saccharomyces cerevisiae ACP2 gene encodes an essential HMG1-like protein.

作者信息

Haggren W, Kolodrubetz D

机构信息

Department of Microbiology, University of Texas Health Science Center, San Antonio 78284.

出版信息

Mol Cell Biol. 1988 Mar;8(3):1282-9. doi: 10.1128/mcb.8.3.1282-1289.1988.

Abstract

The high-mobility-group (HMG) proteins, a group of nonhistone chromatin-associated proteins, have been extensively characterized in higher eucaryotic cells. To test the biological function of an HMG protein, we have cloned and mutagenized a gene encoding an HMG-like protein from the yeast Saccharomyces cerevisiae. A yeast genomic DNA library was screened with an oligonucleotide designed to hybridize to any yeast gene containing an amino acid sequence conserved in several higher eucaryotic HMG proteins. DNA sequencing and Northern (RNA) blot analysis revealed that one gene, called ACP2 (acidic protein 2), synthesizes a poly(A)+ RNA in S. cerevisiae which encodes a 27,000-molecular-weight protein whose amino acid sequence is homologous to those of calf HMG1 and HMG2 and trout HMGT proteins. Standard procedures were used to construct a diploid yeast strain in which one copy of the ACP2 gene was mutated by replacement with the URA3 gene. When this diploid was sporulated and dissected, only half of the spores were viable. About half of the nonviable spores proceeded through two or three cell divisions and then stopped dividing; the rest did not germinate at all. None of the viable spores contained the mutant ACP2 gene, thus proving that the protein encoded by ACP2 is required for cell viability. The results presented here demonstrate that an HMG-like protein has an essential physiological function.

摘要

高迁移率族(HMG)蛋白是一类与非组蛋白染色质相关的蛋白质,在高等真核细胞中已得到广泛研究。为了测试一种HMG蛋白的生物学功能,我们从酿酒酵母中克隆并诱变了一个编码类HMG蛋白的基因。用一种寡核苷酸筛选酵母基因组DNA文库,该寡核苷酸设计用于与任何含有几种高等真核生物HMG蛋白中保守氨基酸序列的酵母基因杂交。DNA测序和Northern(RNA)印迹分析表明,一个名为ACP2(酸性蛋白2)的基因在酿酒酵母中合成一种聚腺苷酸加尾(poly(A)+)RNA,其编码一种分子量为27000的蛋白质,该蛋白质的氨基酸序列与小牛HMG1和HMG2以及鳟鱼HMGT蛋白的氨基酸序列同源。采用标准程序构建了一种二倍体酵母菌株,其中ACP2基因的一个拷贝被URA3基因取代而发生突变。当这种二倍体进行孢子形成和解剖时,只有一半的孢子是存活的。大约一半不能存活的孢子进行了两到三次细胞分裂,然后停止分裂;其余的根本没有发芽。所有存活的孢子都不含有突变的ACP2基因,因此证明ACP2编码的蛋白质是细胞存活所必需的。此处给出的结果表明,一种类HMG蛋白具有重要的生理功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/219a/363274/78ebad244b4c/molcellb00063-0285-a.jpg

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