Evolution of aldolase gene in insect and vertebrate

昆虫和脊椎动物醛缩酶基因的进化

• 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; aldolase; isozyme; Drosophila melanogater; Bombyx mori; gene expression; molecular evolution; invertebrate

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.

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转染到大肠杆菌细胞中，尽管仅部分纯化了同工酶γ，但通过简单的程序将同工酶α和β被纯化为均匀性。同工酶之间的性能相似。新颖的mRNA（命名为4-3和6-2）也来自果蝇成人。 4-3有两个最后的外显子4Alpha和4beta未贴合。发现从4-3个mRNA的产物是主要结构和酶学特性的同工酶α。在D.Melanogaster的组织中，已知编码Exon4Alpha的mRNA的产生被限制为低水平。我们得出的结论是，在RNA处理过程中，可以在外显子4Beta中产生的成绩单编码外显子4Alpha和4beta被识别为poly（a）信号。 6-2克隆具有FBP和FLP的活性位点残基（K*Q）的变体（K*Q）和藻酶活性。这可能具有藻酶的分子进化线索。在Bombyx Mori中，发现并研究了两种类型的醛糖酶同工酶，S和F。器官中五个成分的出现可能是由于S和F亚基的异光原样的形成。ALDOLASE在内部结构中具有四个同工酶特异性组序列。在实验和融合蛋白和果蝇藻酶同工酶CAB中获得的收益如下：（a）IgS-1和4负责确定脊椎动物醛酶异糖酶A和C的组织特异性，以及果蝇藻酶。在其他IGS中，IGS-4可能更换。