Adeno-associated virus type 2 nonstructural protein Rep78 suppresses translation in vitro.
Adeno-associated virus type 2 nonstructural protein Rep78 [621 amino acids (aa) long] affects the expression of various cellular and viral genes. In this study we examined the effects of Rep78 on expression of the luciferase gene from the human cytomegalovirus immediate-early promoter in HeLa cells and on translation of RNA encoding luciferase in rabbit reticulocyte lysate. When Rep78 and luciferase were coexpressed, the luciferase activity decreased despite increased levels of luciferase mRNA in the cells. Purified Rep78 or Rep68 fused with Escherichia coli maltose binding protein suppressed translation of luciferase RNA in vitro, but Rep52/40 fusion proteins did not. A mutated Rep78, which is 520 aa long and truncated at its C-terminus, did suppress the in vitro translation, whereas a similarly truncated Rep78 of 420 aa did not. The results indicate that Rep78/68 function to suppress gene expression through translation inhibition, which requires the N-terminal region contained within aa 1-520.
Takeuchi
T
T
Division of Molecular Genetics, National Institute of Infectious Diseases, 1-23-1, Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
Kozuka
T
T
Nakagawa
K
K
Aoki
Y
Y
Ohtomo
K
K
Yoshiike
K
K
Kanda
T
T
D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D017781:Genes, Immediate-Early; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011817:Rabbits; D011993:Recombinant Fusion Proteins; D012156:Reticulocytes; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014764:Viral Proteins; C045264:replication initiator protein; C068734:rep proteins, Adeno-associated virus 2; D008156:Luciferases; D004265:DNA Helicases