Mason Stephen W, Aberg Erika, Lawetz Carol, DeLong Rachel, Whitehead Paul, Liuzzi Michel
Biological Sciences Department, Boehringer Ingelheim (Canada) Ltd, Laval, Québec, Canada.
J Virol. 2003 Oct;77(19):10670-6. doi: 10.1128/jvi.77.19.10670-10676.2003.
We have investigated protein-protein interactions among the respiratory syncytial virus (RSV) RNA polymerase subunits using affinity chromatography. Here we demonstrate a novel interaction of P and M2-1 proteins. Phosphorylation of either M2-1 or P appears to be dispensable for this interaction. Internal deletions within P mapped the M2-1-binding domain to a region between residues 100 and 120. Alanine-scanning mutagenesis within this region of P revealed that substitution of any one of the three residues, L101, Y102, and F109, prevented both M2-1 and P binding and expression of an M2-1-dependent luciferase reporter gene. However, these same mutations did not prevent the activity of an M2-1-independent chloramphenicol acetyltransferase minigenome, suggesting that these residues of P specifically affect M2-1-P interaction. On the basis of these observations, it is possible that the interaction between RSV M2-1 and P proteins is important for viral replication.
我们利用亲和色谱法研究了呼吸道合胞病毒（RSV）RNA聚合酶亚基之间的蛋白质-蛋白质相互作用。在此，我们展示了P蛋白和M2-1蛋白之间的一种新型相互作用。M2-1或P的磷酸化对于这种相互作用似乎并非必需。P蛋白内部缺失将M2-1结合域定位到第100至120位残基之间的区域。对P蛋白这一区域进行丙氨酸扫描诱变发现，L101、Y102和F109这三个残基中的任何一个被取代，都会阻止M2-1和P的结合以及M2-1依赖性荧光素酶报告基因的表达。然而，这些相同的突变并不影响M2-1非依赖性氯霉素乙酰转移酶微型基因组的活性，这表明P蛋白的这些残基特异性地影响M2-1-P相互作用。基于这些观察结果，RSV M2-1和P蛋白之间的相互作用可能对病毒复制很重要。
