Lysines K117 and K147 play conserved roles in Ras activation from Drosophila to mammals.
Lysines K117 and K147 play conserved roles in Ras activation from Drosophila to mammals.
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DOI：
10.1093/g3journal/jkad201
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发表时间：
2023-11-01
影响因子：
2.6
通讯作者：
Pfleger, Cathie M.
中科院分区：
文献类型：
--
作者：
Singh, Jiya;Karunaraj, Prashath;Luf, Max;Pfleger, Cathie M.
Ras信号传导在生长、增殖和发育模式中起重要作用。维持适当水平的Ras信号传导对于建立发育模式和预防成熟生物体中的疾病（如癌症）非常重要。Ras蛋白在果蝇中以Ras 85 D为代表，在哺乳动物中以HRas、NRas和KRas为代表。在过去的十几年里，许多报道都描述了Ras蛋白的抑制性和激活性泛素化事件。由Rabex-5或Lztr 1介导的抑制性Ras泛素化在果蝇和哺乳动物之间高度保守。在哺乳动物HRas、NRas和KRas中已经报道了K117和K147处的激活泛素化事件，但是尚不清楚K117和K147的这些激活作用是否在果蝇中是保守的。解决这些赖氨酸在果蝇Ras激活中的潜在保守作用需要足够强的表型来评估抑制。因此，我们利用致癌Ras，RasG 12 V，其使Ras偏向于GTP负载的活性构象。我们分别创建了双突变体RasG 12 V、K117 R和RasG 12 V、K147 R以及三突变体RasG 12 V、K117 R、K147 R以防止K117、K147或两者的赖氨酸特异性翻译后修饰。我们将它们的表型与翅膀中的RasG 12 V进行比较，以揭示这些赖氨酸的作用。尽管RasG 12 V、K147 R与RasG 12 V没有显示出令人信服的或可量化的差异，但RasG 12 V、K117 R与RasG 12 V相比显示出可见的和可量化的抑制，并且三重突变体RasG 12 V、K117 R、K147 R与RasG 12 V相比显示出显著的抑制，并且与RasG 12 V、K117 R相比显示出增加的抑制。这些数据与K117和K147在从果蝇到哺乳动物的Ras激活中的高度保守作用一致。
Ras signaling plays an important role in growth, proliferation, and developmental patterning. Maintaining appropriate levels of Ras signaling is important to establish patterning in development and to prevent diseases such as cancer in mature organisms. The Ras protein is represented by Ras85D in Drosophila and by HRas, NRas, and KRas in mammals. In the past dozen years, multiple reports have characterized both inhibitory and activating ubiquitination events regulating Ras proteins. Inhibitory Ras ubiquitination mediated by Rabex-5 or Lztr1 is highly conserved between flies and mammals. Activating ubiquitination events at K117 and K147 have been reported in mammalian HRas, NRas, and KRas, but it is unclear if these activating roles of K117 and K147 are conserved in flies. Addressing a potential conserved role for these lysines in Drosophila Ras activation requires phenotypes strong enough to assess suppression. Therefore, we utilized oncogenic Ras, RasG12V, which biases Ras to the GTP-loaded active conformation. We created double mutants RasG12V,K117R and RasG12V,K147R and triple mutant RasG12V,K117R,K147R to prevent lysine-specific post-translational modification of K117, K147, or both, respectively. We compared their phenotypes to RasG12V in the wing to reveal the roles of these lysines. Although RasG12V,K147R did not show compelling or quantifiable differences from RasG12V, RasG12V,K117R showed visible and quantifiable suppression compared to RasG12V, and triple mutant RasG12V,K117R,K147R showed dramatic suppression compared to RasG12V and increased suppression compared to RasG12V,K117R. These data are consistent with highly conserved roles for K117 and K147 in Ras activation from flies to mammals.
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影响因子：
18.4
作者：
Haigis KM
通讯作者：
Haigis KM
影响因子：
48
作者：
Mertins, Philipp;Qiao, Jana W.;Patel, Jinal;Udeshi, Namrata D.;Clauser, Karl R.;Mani, D. R.;Burgess, Michael W.;Gillette, Michael A.;Jaffe, Jacob D.;Carr, Steven A.
通讯作者：
Carr, Steven A.
影响因子：
2.7
作者：
Guarner, Ana;Manjon, Cristina;Sanchez-Herrero, Ernesto
通讯作者：
Sanchez-Herrero, Ernesto
影响因子：
16.8
作者：
Akimov, Vyacheslav;Barrio-Hernandez, Inigo;Blagoev, Blagoy
通讯作者：
Blagoev, Blagoy
DOI：
10.1074/mcp.m112.017905
发表时间：
2012-12
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Wagner SA;Beli P;Weinert BT;Schölz C;Kelstrup CD;Young C;Nielsen ML;Olsen JV;Brakebusch C;Choudhary C
通讯作者：
Choudhary C