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第二种果蝇蛋白激酶A催化亚基基因的活性、表达及功能

Activity, expression and function of a second Drosophila protein kinase A catalytic subunit gene.

作者信息

Meléndez A, Li W, Kalderon D

机构信息

Department of Biological Sciences, Columbia University, New York, New York 10027, USA.

出版信息

Genetics. 1995 Dec;141(4):1507-20. doi: 10.1093/genetics/141.4.1507.

Abstract

The DC2 gene was isolated previously on the basis of sequence similarity to DC0, the major Drosophila protein kinase A (PKA) catalytic subunit gene. We show here that the 67-kD Drosophila DC2 protein behaves as a PKA catalytic subunit in vitro. DC2 is transcribed in mesodermal anlagen of early embryos. This expression depends on dorsal but on neither twist nor snail activity. DC2 transcriptional fusions mimic this embryonic expression and are also expressed in subsets of cells in the optic lamina, wing disc and leg discs of third instar larvae. A saturation screen of a small deficiency interval containing DC2 for recessive lethal mutations yielded no DC2 alleles. We therefore isolated new deficiencies to generate deficiency trans-heterozygotes that lacked DC2 activity. These animals were viable and fertile. The absence of DC2 did not affect the viability or phenotype of imaginal disc cells lacking DC0 activity or embryonic hatching of animals with reduced DC0 activity. Furthermore, transgenes expressing DC2 from a DC0 promoter did not efficiently rescue a variety of DC0 mutant phenotypes. These observations indicate that DC2 is not an essential gene and is unlikely to be functionally redundant with DC0, which has multiple unique functions during development.

摘要

DC2基因先前是基于与DC0(果蝇主要的蛋白激酶A(PKA)催化亚基基因)的序列相似性而分离得到的。我们在此表明,67-kD的果蝇DC2蛋白在体外表现为PKA催化亚基。DC2在早期胚胎的中胚层原基中被转录。这种表达依赖于背侧,但不依赖于twist或蜗牛的活性。DC2转录融合体模拟这种胚胎表达,并且也在三龄幼虫的视叶、翅芽和腿芽的细胞亚群中表达。对包含DC2的一个小缺失区间进行隐性致死突变的饱和筛选未产生DC2等位基因。因此,我们分离了新的缺失以产生缺乏DC2活性的缺失反式杂合子。这些动物是存活且可育的。DC2的缺失并不影响缺乏DC0活性的成虫盘细胞的活力或表型,也不影响DC0活性降低的动物的胚胎孵化。此外,从DC0启动子表达DC2的转基因不能有效地挽救多种DC0突变表型。这些观察结果表明,DC2不是一个必需基因,并且不太可能与DC0在功能上冗余,DC0在发育过程中具有多种独特功能。

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