Detection of beet yellows closterovirus methyltransferase-like and helicase-like proteins in vivo using monoclonal antibodiesстатья
Статья опубликована в высокорейтинговом журнале
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Дата последнего поиска статьи во внешних источниках: 18 июля 2013 г.
Аннотация:In the positive-stranded RNA genome of beet yellows closterovirus (BYV), the 5'-terminal ORF la encodes a 295 kDa polyprotein with the domains of papain-like cysteine proteinase, methyltransferase (MT) and helicase (HEL), whereas ORF Ib encodes an RNA-dependent RNA polymerase. Eleven and five hybridoma cell lines secreting monoclonal antibodies (MAbs) were derived from mice injected with the bacterially expressed fragments of the BW 1 a product encompassing the MT and HEL domains, respectively, On immunoblots of protein from BW-infected Tetragonia expansa plants, four MAbs against the MT recognized a similar to 63 kDa protein, and two MAbs against the HEL recognized a similar to 100 kDa protein. Both the methyltransferase-like protein and the helicase-like protein were found mainly in the fractions of large organelles (P1) and membranes (P30) of the infected plants. These data clearly indicate that (i) the BW methyltransferase-like and helicase-like proteins, like other related viral enzymes, are associated with membrane compartments in cells, and (ii) the la protein, apart from the cleavage by the leader papain-like proteinase that is expected to produce the 66 kDa and 229 kDa fragments, undergoes additional processing by a virus-encoded or cellular proteinase.