PROTEIN-INTERACTION CLONING IN YEAST - IDENTIFICATION OF MAMMALIAN PROTEINS THAT REACT WITH THE LEUCINE ZIPPER OF JUN
PROTEIN-INTERACTION CLONING IN YEAST - IDENTIFICATION OF MAMMALIAN PROTEINS THAT REACT WITH THE LEUCINE ZIPPER OF JUN
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DOI：
10.1073/pnas.89.13.5789
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发表时间：
1992-07-01
影响因子：
11.1
通讯作者：
NATHANS, D
中科院分区：
文献类型：
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作者：
CHEVRAY, PM;NATHANS, D
为了鉴定与Jun或Fos相互作用的蛋白质，我们使用了由S.菲尔兹和O. K. Song [（1989）Nature（伦敦）340，245-2461克隆编码与二聚化和DNA结合基序相互作用的多肽的哺乳动物cDNA（bZIP;碱性结构域亮氨酸拉链基序）。为此，缺乏GAL 4活性但表达GAL 4 DNA结合结构域的酵母细胞，用小鼠胚胎cDNA质粒文库转化Jun bZIP融合蛋白，其中cDNA连接到编码GAL 4转录激活结构域的基因片段。鉴定了几个表现出GAL 4活性的转化体，并显示其含有编码预测与Jun和/或Fos形成卷曲螺旋结构的多肽的质粒。其中之一是ATF/CREB蛋白家族的bZIP蛋白-可能是TAXREB 67的鼠同源物。另外两个编码多肽的预测潜力，形成卷曲螺旋结构，和其他七个分离物编码的α或β-原肌球蛋白，经典的卷曲螺旋蛋白的片段。发现原肌球蛋白多肽在酵母测定系统中与Jun的bZIP区域相互作用，但不与Fos的bZIP区域相互作用。我们的研究结果说明了蛋白质相互作用克隆的范围，用于发现与给定靶多肽结合的蛋白质。
To identify proteins that interact with Jun or Fos we have used the protein interaction cloning system developed by S. Fields and O.-K. Song [(1989) Nature (London) 340, 245-2461 to clone mammalian cDNAs encoding polypeptides that interact with the dimerization and DNA-binding motif (bZIP; basic domain leucine zipper motif) of Jun. For this purpose, yeast cells lacking GAL4 activity but expressing a GAL4 DNA-binding domain-Jun bZIP fusion protein were transformed with a mouse embryo cDNA plasmid library in which the cDNA was joined to a gene segment encoding the GAL4 transcriptional activation domain. Several transformants exhibiting GAL4 activity were identified and shown to harbor plasmids encoding polypeptides predicted to form coiled-coil structures with Jun and/or Fos. One of these is a bZIP protein of the ATF/CREB protein family-probably the murine homolog of TAXREB67. Two others encode polypeptides with predicted potential to form coiled-coil structures, and seven other isolates encode segments of alpha- or beta-tropomyosin, classical coiled-coil proteins. The tropomyosin polypeptides were found to interact in the yeast assay system with the bZIP region of Jun but not with the bZIP region of Fos. Our results illustrate the range of protein interaction cloning for discovering proteins that bind to a given target polypeptide.