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nrc-1和nrc-2这两个基因的分离与鉴定,这两个基因编码控制粗糙脉孢菌生长和发育的蛋白激酶。

The isolation and characterization of nrc-1 and nrc-2, two genes encoding protein kinases that control growth and development in Neurospora crassa.

作者信息

Kothe G O, Free S J

机构信息

Department of Biological Sciences, State University of New York, Buffalo, New York 14260-1300, USA.

出版信息

Genetics. 1998 May;149(1):117-30. doi: 10.1093/genetics/149.1.117.

Abstract

Using an insertional mutagenesis approach, a series of Neurospora crassa mutants affected in the ability to control entry into the conidiation developmental program were isolated. One such mutant, GTH16-T4, was found to lack normal vegetative hyphae and to undergo constitutive conidiation. The affected gene has been named nrc-1, for nonrepressible conidiation gene #1. The nrc-1 gene was cloned from the mutant genomic DNA by plasmid rescue, and was found to encode a protein closely related to the protein products of the Saccharomyces cerevisiae STE11 and Schizosaccharomyces pombe byr2 genes. Both of these genes encode MAPKK kinases that are necessary for sexual development in these organisms. We conclude the nrc-1 gene encodes a MAPKK kinase that functions to repress the onset of conidiation in N. crassa. A second mutant, GTH16-T17, was found to lack normal vegetative hyphae and to constitutively enter, but not complete, the conidiation program. The affected locus is referred to as nrc-2 (nonrepressible conidiation gene #2). The nrc-2 gene was cloned and found to encode a serine-threonine protein kinase. The kinase is closely related to the predicted protein products of the S. pombe kad5, and the S. cerevisiae YNRO47w and KIN82 genes, three genes that have been identified in genome sequencing projects. The N. crassa nrc-2 gene is the first member of this group of kinases for which a phenotype has been defined. We conclude a functional nrc-2-encoded serine/threonine kinase is required to repress entry into the conidiation program.

摘要

采用插入诱变方法,分离出了一系列在控制进入分生孢子形成发育程序能力方面存在缺陷的粗糙脉孢菌突变体。其中一个这样的突变体GTH16 - T4,被发现缺乏正常的营养菌丝,并进行组成型分生孢子形成。受影响的基因被命名为nrc - 1,即不可抑制的分生孢子形成基因#1。通过质粒拯救从突变体基因组DNA中克隆出nrc - 1基因,发现它编码一种与酿酒酵母STE11和粟酒裂殖酵母byr2基因的蛋白质产物密切相关的蛋白质。这两个基因都编码丝裂原活化蛋白激酶激酶激酶(MAPKK激酶),它们是这些生物体有性发育所必需的。我们得出结论,nrc - 1基因编码一种MAPKK激酶,其功能是抑制粗糙脉孢菌分生孢子形成的起始。另一个突变体GTH16 - T17,被发现缺乏正常的营养菌丝,并组成型进入但未完成分生孢子形成程序。受影响的位点被称为nrc - 2(不可抑制的分生孢子形成基因#2)。克隆出nrc - 2基因,发现它编码一种丝氨酸 - 苏氨酸蛋白激酶。该激酶与粟酒裂殖酵母kad5以及酿酒酵母YNRO47w和KIN82基因的预测蛋白质产物密切相关,这三个基因是在基因组测序项目中鉴定出来的。粗糙脉孢菌nrc - 2基因是这组激酶中第一个被定义了表型的成员。我们得出结论,功能性的nrc - 2编码的丝氨酸/苏氨酸激酶是抑制进入分生孢子形成程序所必需的。

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