Martin Dalziel, Fabio Dall'Olio, Arron Mungul, Véronique Piller, Friedrich Piller
{"title":"Ras癌基因通过ralgef介导的信号向其管家启动子诱导β -半乳糖苷α 2,6-唾液转移酶(ST6Gal I)。","authors":"Martin Dalziel, Fabio Dall'Olio, Arron Mungul, Véronique Piller, Friedrich Piller","doi":"10.1111/j.1432-1033.2004.04284.x","DOIUrl":null,"url":null,"abstract":"<p><p>Several oncogenic proteins are known to influence cellular glycosylation. In particular, transfection of codon 12 point mutated H-Ras increases CMP-Neu5Ac: Galbeta1,4GlcNAc alpha2,6-sialyltransferase I (ST6Gal I) activity in rodent fibroblasts. Given that Ras mediates its effects through at least three secondary effector pathways (Raf, RalGEFs and PI3K) and that transcriptional control of mouse ST6Gal I is achieved by the selective use of multiple promoters, we attempted to identify which of these parameters are involved in linking the Ras signal to ST6Gal I gene transcription in mouse fibroblasts. Transformation by human K-Ras or H-Ras (S12 and V12 point mutations, respectively) results in a 10-fold increase in ST6Gal I mRNA, but no alteration in the expression of related sialyltransferases. Using an inducible H-RasV12 expression system, a direct causal link between activated H-Ras expression and elevated ST6Gal I mRNA was demonstrated. The accumulation of the ST6Gal I transcript in response to activated Ras was accompanied by an increase of alpha2,6-sialyltransferase activity and of Neu5Acalpha2,6Gal at the cell surface. Results obtained with H-RasV12 partial loss of function mutants H-RasV12S35 (Raf signal only), H-RasV12C40 (PI3-kinase signal only) and H-RasV12G37 (RalGEFs signal only) suggest that the H-Ras induction of the mouse ST6Gal I gene (Siat1) transcription is primarily routed through RalGEFs. 5'-Rapid amplification of cDNA ends analysis demonstrated that the increase in ST6Gal I mRNA upon H-RasV12 or K-RasS12 transfection is mediated by the Siat1 housekeeping promoter P3-associated 5' untranslated exons.</p>","PeriodicalId":11817,"journal":{"name":"European journal of biochemistry","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2004-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/j.1432-1033.2004.04284.x","citationCount":"52","resultStr":"{\"title\":\"Ras oncogene induces beta-galactoside alpha2,6-sialyltransferase (ST6Gal I) via a RalGEF-mediated signal to its housekeeping promoter.\",\"authors\":\"Martin Dalziel, Fabio Dall'Olio, Arron Mungul, Véronique Piller, Friedrich Piller\",\"doi\":\"10.1111/j.1432-1033.2004.04284.x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Several oncogenic proteins are known to influence cellular glycosylation. 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引用次数: 52
摘要
已知几种致癌蛋白影响细胞糖基化。特别是,转染密码子12点突变的H-Ras增加了啮齿动物成纤维细胞中CMP-Neu5Ac: Galbeta1,4GlcNAc alpha2,6-唾液转移酶I (ST6Gal I)的活性。考虑到Ras通过至少三种次级效应通路(Raf、ralgef和PI3K)介导其作用,以及小鼠ST6Gal I的转录控制是通过选择性使用多个启动子实现的,我们试图确定这些参数中哪些参与了Ras信号与小鼠成纤维细胞中ST6Gal I基因转录的联系。人K-Ras或H-Ras(分别为S12和V12点突变)的转化导致ST6Gal I mRNA增加10倍,但相关唾液转移酶的表达没有改变。通过诱导H-RasV12表达系统,证实了H-Ras表达激活与ST6Gal I mRNA升高之间的直接因果关系。ST6Gal I转录物在Ras激活下的积累伴随着细胞表面α - 2,6-唾液基转移酶活性和neu5acα - 2,6gal活性的增加。H-RasV12部分功能缺失突变体H-RasV12S35(仅Raf信号)、H-RasV12C40(仅pi3激酶信号)和H-RasV12G37(仅RalGEFs信号)的结果表明,H-Ras诱导小鼠ST6Gal I基因(Siat1)转录主要通过RalGEFs进行。5'- cDNA末端快速扩增分析表明,H-RasV12或K-RasS12转染后ST6Gal I mRNA的增加是由Siat1管家启动子p3相关的5'未翻译外显子介导的。
Ras oncogene induces beta-galactoside alpha2,6-sialyltransferase (ST6Gal I) via a RalGEF-mediated signal to its housekeeping promoter.
Several oncogenic proteins are known to influence cellular glycosylation. In particular, transfection of codon 12 point mutated H-Ras increases CMP-Neu5Ac: Galbeta1,4GlcNAc alpha2,6-sialyltransferase I (ST6Gal I) activity in rodent fibroblasts. Given that Ras mediates its effects through at least three secondary effector pathways (Raf, RalGEFs and PI3K) and that transcriptional control of mouse ST6Gal I is achieved by the selective use of multiple promoters, we attempted to identify which of these parameters are involved in linking the Ras signal to ST6Gal I gene transcription in mouse fibroblasts. Transformation by human K-Ras or H-Ras (S12 and V12 point mutations, respectively) results in a 10-fold increase in ST6Gal I mRNA, but no alteration in the expression of related sialyltransferases. Using an inducible H-RasV12 expression system, a direct causal link between activated H-Ras expression and elevated ST6Gal I mRNA was demonstrated. The accumulation of the ST6Gal I transcript in response to activated Ras was accompanied by an increase of alpha2,6-sialyltransferase activity and of Neu5Acalpha2,6Gal at the cell surface. Results obtained with H-RasV12 partial loss of function mutants H-RasV12S35 (Raf signal only), H-RasV12C40 (PI3-kinase signal only) and H-RasV12G37 (RalGEFs signal only) suggest that the H-Ras induction of the mouse ST6Gal I gene (Siat1) transcription is primarily routed through RalGEFs. 5'-Rapid amplification of cDNA ends analysis demonstrated that the increase in ST6Gal I mRNA upon H-RasV12 or K-RasS12 transfection is mediated by the Siat1 housekeeping promoter P3-associated 5' untranslated exons.