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Microbiology 152 (2006), 3467-3476; DOI  10.1099/mic.0.29134-0
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Microbiology 152 (2006), 3467-3476; DOI  10.1099/mic.0.29134-0
© 2006 Society for General Microbiology

tRNA-dependent cleavage of the ColE1 plasmid-encoded RNA I

Zhijun Wang1,2, Zhenghong Yuan1, Li Xiang1, Junjie Shao1 and Grzegorz Wegrzyn3,4

1 Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 200032, Shanghai, People's Republic of China
2 Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca 14853, NY, USA
3 Department of Molecular Biology, University of Gdansk, 80-822, Gdansk, Poland
4 Department of Genetics and Marine Biotechnology, Institute of Oceanology, Polish Academy of Sciences, Sw. Wojciecha 5, 81-347 Gdynia, Poland

Correspondence
Grzegorz Wegrzyn
wegrzyn{at}biotech.univ.gda.pl

Effects of tRNAAla(UGC) and its derivative devoid of the 3'-ACCA motif [tRNAAla(UGC){Delta}ACCA] 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 tRNAAla(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 tRNAAla(UGC). Moreover, tRNAAla(UGC), but not tRNAAla(UGC){Delta}ACCA, caused RNA I cleavage in vitro in the presence of MgCl2. These results strongly suggest that tRNA-dependent RNA I cleavage occurs in ColE1-like plasmid-bearing E. coli, and demonstrate that tRNAAla(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 tRNAAla(UGC).






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