Hepatitis B virus (HBV), a major global health problem, can cause chronic hepatitis, liver cirrhosis, and hepatocellular carcinomas in chronically infected patients. However, before HBV infection can be adequately controlled, many mysteries about the HBV life cycle must be solved. In this study, TIMM29, an inner mitochondrial membrane protein, was identified as an interaction partner of the preS1 region of the HBV large S protein. The interaction was verified by both an immunoprecipitation with preS1 peptides and a GST‐pulldown assay. Immunofluorescence studies also showed colocalization of preS1 and TIMM29. Moreover, it was determined that the preS1 bound with amino acids 92–189 of the TIMM29 protein. Infection of HBV in TIMM29‐overexpressing NTCP/G2 cells resulted in a significant decrease of HBeAg and both extracellular particle‐associated and core particle‐associated HBV DNA without affecting cccDNA formation. Comparable results were obtained with TIMM29‐overexpressing HB611 cells, which constitutively produce HBV. In contrast, knockout of TIMM29 in NTCP/G2 cells led to a higher production of HBV including HBeAg expression, as did knockout of TIMM29 in HB611. Collectively, these results suggested that TIMM29 interacts with the preS1 region of the HBV large S protein and modulates HBV amplification.

中文翻译：

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TIMM29与乙型肝炎病毒preS1相互作用以调节HBV生命周期。

乙型肝炎病毒（HBV）是一个主要的全球性健康问题，可在慢性感染的患者中引起慢性肝炎，肝硬化和肝细胞癌。但是，在可以充分控制HBV感染之前，必须解决许多有关HBV生命周期的谜团。在这项研究中，内部线粒体膜蛋白TIMM29被确定为HBV大S蛋白preS1区域的相互作用伴侣。通过preS1肽的免疫沉淀和GST-pulldown试验验证了相互作用。免疫荧光研究还显示了preS1和TIMM29的共定位。此外，已确定preS1与TIMM29蛋白的92-189位氨基酸结合。在过表达TIMM29的NTCP / G2细胞中感染HBV导致HBeAg以及细胞外颗粒相关的HBV DNA和核心颗粒相关的HBV DNA显着降低，而不会影响cccDNA的形成。使用TIMM29过表达的HB611细胞（可组成性产生HBV）获得了可比的结果。相反，敲除NTCP / G2细胞中的TIMM29会导致产生更高的HBV，包括HBeAg表达，就像敲除HB611中的TIMM29一样。总体而言，这些结果表明TIMM29与HBV大S蛋白的preS1区域相互作用并调节HBV扩增。NTCP / G2细胞中TIMM29的敲除导致HBV的更高产量，包括HBeAg表达，而HB611中TIMM29的敲除也是如此。总体而言，这些结果表明TIMM29与HBV大S蛋白的preS1区域相互作用并调节HBV扩增。NTCP / G2细胞中TIMM29的敲除导致HBV的更高产量，包括HBeAg表达，而HB611中TIMM29的敲除也是如此。总体而言，这些结果表明TIMM29与HBV大S蛋白的preS1区域相互作用并调节HBV扩增。