EVOLUTIONARY STUDY OF ENVIRONMENT-DEPENDENT MUTATIONS OF TRIMERESURUS FLAVOVIRIDIS VENOM-GLAND PHOSPHOLIPASE A2 ISOZYME GENES

竹叶青毒腺磷脂酶A2同工酶基因环境依赖性突变的进化研究

• 批准号: 12640689
• 负责人: OHNO Ryuichi
• 金额: $ 2.3万
• 依托单位: SOJO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12640689/
• 关键词: Southwestem islands of Japan; Trimeresurus flavoviridis; Isolated environments; Venom-gland phospholipase A2 isozymes; Gene mutation; Pseudogene; Feeding habit; Adaptive evolution; ハブ毒; クロマトグラフィー; ホスホリパービA_2アイソザイム; 孤立

Trimeresurus flavoviridis (Tf) snakes inhabit the southwestern islands of Japan : Amami-Oshima, Tokunoshima and Okinawa. These islands are thought to have been separated one to two million years ago by eustacy (changes in sea level) in the erogenic stage. This work is to correlate mutations of Tf venom-gland phospholipase A_2 (PLA_2) isozyme genes with their isolated environments. When the venoms of Amami-Oshima, Tokunoshima and Okinawa Tf snakes were chromatographed, two [Lys^<49>]PLA_2s called BPI and BPII are missing only in Okinawa Tf venom. Northern blot analysis showed the absence of BPI and BPII mRNAs in Okinawa Tf venom gland. Amplification by polymerase chain reaction (PCR) of the segments of BPI and BPII genes with specific primers against Okinawa Tf genomic DNA together with PCR amplification of the segments including polymorphic positions in the upstream and downstream regions of BPI and BPII genes and analysis of their sequences established that the upstream region of BPI … More gene and the downstream region of BPII gene are fused to form pseudogene in Okinawa Tfgenome. Since BPI and BPII have strong necrotic activity, it is thought that the lack of BPI and BPII is of great disadvantage to Okinawa Tf. However, they seem to subsist well. In Yambaru, a mountain area covered with forests in the northern part of Okinawa, Tf snakes mostly (90%) prey on Hoist's frogs (Rana holsti) inhabiting the streams in this area. This natural, wild area seems to reflect the environment in Okinawa in which Tfsnakes lived in ancient times. It could be assumed that these feeding habits over a long period affected the venom components in Okinawa Tf and the lack of necessity for a strong venom toxicity gave rise to inactivation of BPI and BPII genes in an adaptive manner. PLA-B from Tokunoshima Tf venom can produce edema. However, PL-Y in Okinawa Tf venom which has only three substituted residues, is inactive for edema. Such mutation is also dependent on the difference in their living environments. Less

黄龙蛇生活在日本西南部的三个岛屿：大岛、德岛和冲绳。这些岛屿被认为是一两百万年前在造山期因海平面变化而分开的。本研究旨在探讨转移因子毒腺磷脂酶A_2(PLA2)同工酶基因突变与分离环境的关系。当对Amami-Oshima、Tokunoshima和Okinawa TF蛇毒进行层析时，只有冲绳TF蛇毒中缺失了两个名为BPI和BPII的PLA2。Northern印迹分析表明，冲绳TF毒腺中没有BPI和BPII基因。用针对冲绳Tf基因组DNA的特异性引物对bpi和bpII基因片段进行聚合酶链式反应扩增，并对含有bpI和bpII基因上下游多态位置的片段进行聚合酶链式反应扩增，并对其序列进行分析，证实bpI…的上游区域在冲绳Tf基因组中，更多的基因与BPII基因下游区域融合形成假基因。由于BPI和BPII具有很强的坏死活性，因此认为缺乏BPI和BPII对冲绳TF非常不利。然而，他们似乎生存得很好。在冲绳北部森林覆盖的山区Yambaru，TF蛇主要(90%)捕食栖息在该地区溪流中的Hoist‘s青蛙(Rana Holsti)。这片自然、荒野的区域似乎反映了冲绳古代蛇类的生存环境。可以推测，这些摄食习性在很长一段时间内影响了冲绳Tf的毒液成分，由于缺乏强烈的毒力，导致BPI和BPII基因以一种适应性的方式失活。德岛TF毒液中的聚乳酸-B可引起水肿症。然而，冲绳TF蛇毒中只有三个取代残基的PL-Y对水肿没有活性。这种突变还取决于它们生活环境的不同。较少

项目成果

Takahito Chijiwa: "Regional evolution of venom-gland phospholipase A_2 isoenzymes of Trimeresurus flavoviridis snakes in the southwestern islands of Japan"Biochemical Journal. 347. 491-499 (2000)

Takahito Chijiwa：“日本西南部岛屿竹叶青蛇毒腺磷脂酶A_2同工酶的区域进化”生物化学杂志。

Yamamoto,C., Ohno,M. et al.: "Trimeresurus flavoviridis (Habu Snake) venom induces human erythrocyte lysis through enzymatic lipolysis, complement activation and decreased membrane expression of CD55 and CD59"Pharmacoloty & Taxicology. 89. 188-194 (2001)

山本，C.，大野，M.

Ohno,M., Chijiwa,T. et al.: "Accelerated and regional evolution of snake venom gland isozymes"Perspectives in Molecular Toxinology (Menez, A., ed.) John Wiley & Sons, London. 387-400 (2002)

大野，M.，Chijiwa，T.

Yoko Yamaguci: "Characterization, amino acid sequence and evolution of edema-inducing, basic phospholipase A_2 from Trimeresurus flavoviridis venom"Toxicon. 39. 1069-1076 (2001)

Yoko Yamaguci：“来自竹叶青蛇毒液的水肿诱导碱性磷脂酶 A_2 的特征、氨基酸序列和进化”Toxicon。

Yoshiro Chuman: "Regional and accelerated molecular evolution in group I snake venom gland phospholipase A_2 isozymes"Toxicon. 38. 449-462 (2000)

Yoshiro Chuman：“I 组蛇毒腺磷脂酶 A_2 同工酶的区域和加速分子进化”Toxicon。