tRNA-dependent cleavage of the ColE1 plasmid-encoded RNA I.
Effects of tRNA(Ala)(UGC) and its derivative devoid of the 3'-ACCA motif [tRNA(Ala)(UGC)DeltaACCA] on the cleavage of the ColE1-like plasmid-derived RNA I were analysed in vivo and in vitro. In an amino-acid-starved relA mutant, in which uncharged tRNAs occur in large amounts, three products of specific cleavage of RNA I were observed, in contrast to an otherwise isogenic relA(+) host. Overexpression of tRNA(Ala)(UGC), which under such conditions occurs in Escherichia coli mostly in an uncharged form, induced RNA I cleavage and resulted in an increase in ColE1-like plasmid DNA copy number. Such effects were not observed during overexpression of the 3'-ACCA-truncated tRNA(Ala)(UGC). Moreover, tRNA(Ala)(UGC), but not tRNA(Ala)(UGC)DeltaACCA, caused RNA I cleavage in vitro in the presence of MgCl(2). These results strongly suggest that tRNA-dependent RNA I cleavage occurs in ColE1-like plasmid-bearing E. coli, and demonstrate that tRNA(Ala)(UGC) participates in specific degradation of RNA I in vivo and in vitro. This reaction is dependent on the presence of the 3'-ACCA motif of tRNA(Ala)(UGC).
Wang
Zhijun
Z
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca 14853, NY, USA.|Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 200032, Shanghai, People's Republic of China.
Yuan
Zhenghong
Z
Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 200032, Shanghai, People's Republic of China.
Xiang
Li
L
Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 200032, Shanghai, People's Republic of China.
Shao
Junjie
J
Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 200032, Shanghai, People's Republic of China.
Węgrzyn
Grzegorz
G
Department of Genetics and Marine Biotechnology, Institute of Oceanology, Polish Academy of Sciences, Św. Wojciecha 5, 81-347 Gdynia, Poland.|Department of Molecular Biology, University of Gdańsk, 80-822, Gdańsk, Poland.
D003086:Bacteriocin Plasmids / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D003067:Coenzymes / Q000494:pharmacology; D004269:DNA, Bacterial / Q000096:biosynthesis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015636:Magnesium Chloride / Q000494:pharmacology; D008954:Models, Biological; D008969:Molecular Sequence Data; D020871:RNA Stability; D016372:RNA, Antisense / Q000235:genetics / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D016337:RNA, Catalytic / Q000378:metabolism*; D012344:RNA, Transfer, Ala / Q000378:metabolism*
D003067:Coenzymes; D004269:DNA, Bacterial; C042027:RNA I; D016372:RNA, Antisense; D012329:RNA, Bacterial; D016337:RNA, Catalytic; D012344:RNA, Transfer, Ala; D015636:Magnesium Chloride