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Evolution of aldolase gene in insect and vertebrate

昆虫和脊椎动物醛缩酶基因的进化

基本信息

批准号：
07660444
负责人：
SUGIMOTO Yasushi
金额：
$ 1.41万
依托单位：
Kagoshima University (1996)Seinan Jo Gakuin University Junior College (1995)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660444/
关键词：
aldolase isozyme Drosophila melanogater Bombyx mori gene expression molecular evolution invertebrate

项目摘要

Fructose-1,6-bisphoshate aldolase is a member of alpha / beta barrel enzyme family and has three types of the isozymes form : A,B and C.Three isozymic forms, alpha, beta and gamma, of Drosophila melanogaster aldolase are produced from a single gene by alternative usage of the triple exons 4. The expression plasmids for the respective isozymes were transfected into Escherichia coli cells, and the isozymes alpha and beta were purified to homogeneity by a simple procedure, though isozyme gamma was only partially purified. The properties were similar among isozymes. The novel-type mRNAs (named, 4-3 and 6-2) was also obtained from Drosophila adult. The 4-3 has two final exons 4alpha and 4beta unspliced. The product from 4-3 mRNA was found to be isozyme alpha from the primary structure and the enzymological properties. In tissues of D.melanogaster, the production of mRNA encoding exon4alpha is known to be restrained to a low level. We concluded that the transcript-encoding exons 4alpha and 4beta, might be produced the coding frame in exon 4beta, is recognized as poly (A) signal during RNA processing. The 6-2 clone has a variant of active site residue (K*Q) and aldolase activity for FBP and FlP.This might have a clue of molecular evolution for aldolase. In Bombyx mori, two types of aldolase isozymes, S and F,were found and studied in emzymology and tissue expressions. The occurrence of five components in the organ may be due to the formation of heterotetramers of the S and F subunits.Aldolase has four isozyme specific group sequences in internal structure. The obtained in the experiments and fusion proteins and Drosophila aldolase isozymes cab be summarised as follows : (a) IGS-1 and 4 are responsible for determining tissues-specificity of vertebrate aldolase isozyme A and C,and Drosophila aldolase. IGS-4 might be more changeable among other IGSs.

果糖-1,6-二磷酸醛缩酶是α / β桶酶家族的一员，有三种同工酶形式：a、B和c。果蝇醛缩酶的α、β和γ三种同工酶形式是由单个基因通过三外显子4的替代使用产生的。将各自同工酶的表达质粒转染到大肠杆菌细胞中，通过简单的程序纯化α和β同工酶至均质，而γ同工酶仅部分纯化。同工酶的性质相似。新型mrna（命名为，4-3和6-2）也从成年果蝇中获得。4-3有两个未拼接的最后外显子4α和4β。从初级结构和酶学性质上确定4-3 mRNA的产物为α同工酶。在黑腹d.m anogaster的组织中，编码外显子4 α的mRNA的产生被限制在低水平。我们认为编码转录的外显子4alpha和4beta可能在RNA加工过程中产生编码框，在外显子4beta中被识别为poly (A)信号。6-2克隆具有FBP和FlP活性位点残基（K*Q）和醛缩酶活性的变异。这可能为醛缩酶的分子进化提供了线索。在家蚕中发现了两种醛缩酶同工酶S和F，并对其酶学和组织表达进行了研究。五组分在器官中的出现可能是由于S和F亚基的异四聚体的形成。醛缩酶在内部结构上具有4个同工酶特异性基团序列。实验中获得的融合蛋白和果蝇醛缩酶同工酶可以总结如下：(a) IGS-1和4负责确定脊椎动物醛缩酶同工酶a和C以及果蝇醛缩酶的组织特异性。IGS-4在其他igs中可能更加多变。