Methionine Mistranslation Bypasses the Restraint of the Genetic Code to Generate Mutant Proteins with Distinct Activities.
Methionine Mistranslation Bypasses the Restraint of the Genetic Code to Generate Mutant Proteins with Distinct Activities.
复制标题
DOI：
10.1371/journal.pgen.1005745
复制
发表时间：
2015-12
期刊：
影响因子：
4.5
通讯作者：
Pan T
中科院分区：
文献类型：
--
作者：
Wang X;Pan T
Although mistranslation is commonly believed to be deleterious, recent evidence indicates that mistranslation can be actively regulated and be beneficial in stress response. Methionine mistranslation in mammalian cells is regulated by reactive oxygen species where cells deliberately alter the proteome through incorporating Met at non-Met positions to enhance oxidative stress response. However, it was not known whether specific, mistranslated mutant proteins have distinct activities from the wild-type protein whose sequence is restrained by the genetic code. Here, we show that Met mistranslation with and without Ca2+ overload generates specific mutant Ca2+/calmodulin-dependent protein kinase II (CaMKII) proteins substituting non-Met with Met at multiple locations. Compared to the genetically encoded wild-type CaMKII, specific mutant CaMKIIs can have distinct activation profiles, intracellular localization and enhanced phenotypes. Our results demonstrate that Met-mistranslation, or “Met-scan” can indeed generate mutant proteins in cells that expand the activity profile of the wild-type protein, and provide a molecular mechanism for the role of regulated mistranslation. Methionine-mistranslation is a recently discovered phenomenon where mammalian cells deliberately mischarge non-Met-tRNAs with amino acid methionine in unstressed cells and in response to innate immune and chemically triggered oxidative stress. These mischarged tRNAs are used in translation to generate mutant proteins containing non-Met to Met substitutions. Accumulating evidence shows that cells employ regulated mistranslation to enhance response to oxidative and other stresses. However, it was unknown whether any specific mutant proteins generated in mistranslation truly have distinct activities as the wild-type protein. Here, we identify and characterize naturally occurring Met-mistranslated proteins in human cells and show that specific Met-mistranslated proteins can have very distinct properties compared to the wild-type protein in vitro and in vivo. For the Ca2+/calmodulin-dependent protein kinase II (CaMKII), Met-mistranslated CaMKII show significant differences in catalytic activities, subcellular localization and elevated caspase-3 activity from the wild-type CaMKII. Our results provide a molecular mechanism on how mistranslation is used as an adaptive mechanism to stress, and establish Met-mistranslation as a nature’s way to overcome the restraint on the protein sequence by the genetic code.
