Smith Alexander C, Cronan John E
Department of Microbiology, University of Illinois, Urbana, Illinois, USA.
Department of Microbiology, University of Illinois, Urbana, Illinois, USA Department of Biochemistry, University of Illinois, Urbana, Illinois, USA
J Bacteriol. 2014 Nov;196(21):3768-75. doi: 10.1128/JB.02091-14. Epub 2014 Aug 25.
In Escherichia coli, synthesis of the malonyl coenzyme A (malonyl-CoA) required for membrane lipid synthesis is catalyzed by acetyl-CoA carboxylase, a large complex composed of four subunits. The subunit proteins are needed in a defined stoichiometry, and it remains unclear how such production is achieved since the proteins are encoded at three different loci. Meades and coworkers (G. Meades, Jr., B. K. Benson, A. Grove, and G. L. Waldrop, Nucleic Acids Res. 38:1217-1227, 2010, doi:http://dx.doi.org/10.1093/nar/gkp1079) reported that coordinated production of the AccA and AccD subunits is due to a translational repression mechanism exerted by the proteins themselves. The AccA and AccD subunits form the carboxyltransferase (CT) heterotetramer that catalyzes the second partial reaction of acetyl-CoA carboxylase. Meades et al. reported that CT tetramers bind the central portions of the accA and accD mRNAs and block their translation in vitro. However, long mRNA molecules (500 to 600 bases) were required for CT binding, but such long mRNA molecules devoid of ribosomes seemed unlikely to exist in vivo. This, plus problematical aspects of the data reported by Meades and coworkers, led us to perform in vivo experiments to test CT tetramer-mediated translational repression of the accA and accD mRNAs. We report that increased levels of CT tetramer have no detectable effect on translation of the CT subunit mRNAs.
在大肠杆菌中,膜脂合成所需的丙二酰辅酶A(丙二酰 - CoA)的合成由乙酰辅酶A羧化酶催化,该酶是一种由四个亚基组成的大型复合物。亚基蛋白需要以特定的化学计量比存在,由于这些蛋白在三个不同的基因座编码,目前尚不清楚这种生产是如何实现的。米兹和同事们(小G. 米兹、B. K. 本森、A. 格罗夫和G. L. 沃尔德罗普,《核酸研究》38:1217 - 1227,2010,doi:http://dx.doi.org/10.1093/nar/gkp1079)报道,AccA和AccD亚基的协同产生是由于蛋白质自身施加的翻译抑制机制。AccA和AccD亚基形成催化乙酰辅酶A羧化酶第二个部分反应的羧基转移酶(CT)异源四聚体。米兹等人报道,CT四聚体结合accA和accD mRNA的中央部分并在体外阻断它们的翻译。然而,CT结合需要长mRNA分子(500至600个碱基),但这种不含核糖体的长mRNA分子似乎不太可能在体内存在。这一点,再加上米兹及其同事报道的数据存在问题,促使我们进行体内实验,以测试CT四聚体介导的accA和accD mRNA的翻译抑制作用。我们报道,CT四聚体水平的增加对CT亚基mRNA的翻译没有可检测到的影响。