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对果蝇中一个影响求偶和视觉介导行为的基因所编码蛋白质中的假定RNA结合基序进行生物测定:非A基因的体外诱变

Bioassaying putative RNA-binding motifs in a protein encoded by a gene that influences courtship and visually mediated behavior in Drosophila: in vitro mutagenesis of nonA.

作者信息

Stanewsky R, Fry T A, Reim I, Saumweber H, Hall J C

机构信息

Department of Biology, Brandeis University, Waltham, Massachusetts 02254, USA.

出版信息

Genetics. 1996 May;143(1):259-75. doi: 10.1093/genetics/143.1.259.

Abstract

The no-on-transient-A (nonA) gene of Drosophila melanogaster influences vision, courtship song, and viability. The nonA-encoded polypeptide is inferred to bind single-stranded nucleic acids. Although sequence-analysis of NONA implies that it belongs to a special interspecific family of this protein type, it does contain two classical RNA recognition motifs (RRM). Their behavioral significance was assayed by generating transgenic strains that were singly or multiply mutated within the relatively N-terminal motif (RRM1) or within RRM2. Neither class of mutation affected NONA binding to polytene chromosomes. The former mutations led to extremely low viability, accompanied by diminished adult longevities that were much worse than for a nonA-null mutant, implying that faulty interpolypeptide interactions might accompany the effects of the amino-acid substitutions within RRM1. All in vitro-mutated types caused optomotor blindness and an absence of transient spikes in the electroretinogram. Courtship analysis discriminated between the effects of the mutations: the RRM2-mutated type generated song pulses and trains that tended to be mildly mutant. These phenotypic abnormalities reinforce the notion that nonA's ubiquitous expression has its most important consequences in the optic lobes, the thoracic ganglia, or both, depending in part on the nonA allele.

摘要

黑腹果蝇的无瞬时A(nonA)基因影响视觉、求偶歌和生存能力。推测由nonA编码的多肽可结合单链核酸。尽管NONA的序列分析表明它属于该蛋白类型的一个特殊种间家族,但它确实包含两个经典的RNA识别基序(RRM)。通过构建在相对N端基序(RRM1)或RRM2内单个或多个发生突变的转基因品系,对它们的行为意义进行了测定。这两类突变均未影响NONA与多线染色体的结合。前一类突变导致极低的生存能力,同时成虫寿命缩短,比nonA基因缺失突变体的情况还要糟糕得多,这意味着RRM1内氨基酸取代的影响可能伴随着多肽间相互作用的缺陷。所有体外突变类型均导致视动性眼震失明和视网膜电图中无瞬时尖峰。求偶分析区分了突变的影响:RRM2突变型产生的歌声脉冲和序列往往有轻度突变。这些表型异常强化了这样一种观念,即nonA的普遍表达在视叶、胸神经节或两者中产生其最重要的影响,部分取决于nonA等位基因。

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