TGMV replication protein AL1 preferentially binds to single‐stranded DNA from the common region

FEBS Letters

Hayes, R.J.¹  Osman, T.A.M.¹  Thömmes, P.¹  Buck, K.W.¹ 

[1] Department of Biology, Imperial College of Science, Technology and Medicine, London SW7 2BB, UK

关键词: Geminivirus;    Replication;    DNA binding protein;    Common region;    Origin of rolling circle replication;    BSA;    bovine serum albumin;    DTT;    dithiothreitol;    ORF;    open reading frame;    PCR;    polymerase chain reaction;    PMSF;    phenylmethylsulfonyl fluoride;    RCNMV;    red clover necrotic mosaic virus;    RCR;    rolling circle replication initiator protein;    TGMV;    tomato golden mosaic virus;   

DOI  :  10.1016/0014-5793(93)80044-U

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The AL1 protein of tomato golden mosaic virus (TGMV) is encoded by the viral DNA and has been shown to be essential for viral DNA replication. We have over-expressed the AL1 open reading frame in E. coli and purified the protein from bacterial extracts to near homogeneity. Using various different techniques we have studied the interaction of the AL1 protein with DNA. The AL1 protein is able to bind to DNA containing the common region of the viral genome, which can be demonstrated by photochemical cross-linking. Binding is 4-fold stronger to single-stranded than to double-stranded DNA. Antibodies against the AL1 protein can be used to precipitate the protein-DNA complex. The binding to single- and double-stranded DNA is specifically to the common region since a DNA fragment unrelated to TGMV is not shifted in a gel retardation assay.

【 授权许可】

Unknown