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第85卷，问题6

呼吸合胞病毒P-P和P-N蛋白复合物的生化特征及P蛋白寡聚化结构域的定位自由

Abstract

依赖RNA的RNA聚合酶和复杂atory syncytial virus (RSV) is composed of the large polymerase (L), the phosphoprotein (P), the nucleocapsid protein (N) and the co-factors M2-1 and M2-2. The P protein plays a central role within the replicase–transcriptase machinery, forming homo-oligomers and complexes with N and L. In order to study P–P and N–P complexes, and the role of P phosphorylation in these interactions, the human RSV P and N proteins were expressed inas His-tagged or GST-fusion proteins. The non-phosphorylated status of recombinant P protein was established by mass spectrometry. GST-P and GST-N fusion proteins were able to interact with RSV proteins extracted from infected cells in a GST pull-down assay. When co-expressed in bacteria, GST-P and His-P were co-purified by glutathione-Sepharose affinity, showing that the RSV P protein can form oligomers within bacteria. This result was confirmed by chemical cross-linking experiments and gel filtration studies. The P oligomerization domain was investigated by a GST pull-down assay using a series of P deletion constructs. This domain was mapped to a small region situated in the central part of P (aa 120–150), which localized in a computer-predicted coiled-coil domain. When co-expressed in bacteria, RSV N and P proteins formed a soluble complex that prevented non-specific binding of N to bacterial RNA. Therefore, RSV P protein phosphorylation is not required for the formation of P–P and N–P complexes, and P controls the RNA binding activity of N.

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2004-06-01.
2022-03-13
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呼吸合胞病毒P-P和P-N蛋白复合物的生化特征及P蛋白寡聚化结构域的定位
J. Gen.Virol。85 ，1643 (2004) ;https://doi.org/10.1099/vir.0.79830-0
/ content/journal/jgv/10.1099/vir.0.79830-0.
/ content/journal/jgv/10.1099/vir.0.79830-0.
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