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The Terminal and Internal Hairpin Loops of the ctRNA of Plasmid pJB01 Play Critical Roles in Regulating Copy Number  

Kim, Sam Woong (Division of Biological Sciences, Pusan National University)
Jeong, In Sil (Division of Applied Life Sciences (Brain Korea 21 and EBNCRC), Graduate School of Gyeongsang National University)
Jeong, Eun Ju (Division of Applied Life Sciences (Brain Korea 21 and EBNCRC), Graduate School of Gyeongsang National University)
Tak, Je Il (Division of Applied Life Sciences (Brain Korea 21 and EBNCRC), Graduate School of Gyeongsang National University)
Lee, John Hwa (College of Veterinary Medicine, Chonbuk National University)
Eo, Seong Kug (College of Veterinary Medicine, Chonbuk National University)
Kang, Ho Young (Division of Biological Sciences, Pusan National University)
Bahk, Jeong Dong (Division of Applied Life Sciences (Brain Korea 21 and EBNCRC), Graduate School of Gyeongsang National University)
Publication Information
Molecules and Cells / v.26, no.1, 2008 , pp. 26-33 More about this Journal
Abstract
The plasmid pJB01, a member of the pMV158 family isolated from Enterococcus faecium JC1, contains three open reading frames, copA, repB, and repC. Plasmids included in this family produce counter-transcribed RNA (ctRNA) that contributes to copy number control. The pJB01 ctRNA, a transcript which consists of 54 nucleotides (nts), is encoded on the opposite strand from the copA/repB intergenic region and partially overlaps an atypical ribosome binding site (ARBS) for repB. The ARBS is integrated by the two underlined conserved regions: 5'-TTTTTGTNNNNTAANNNNNNNNNATG-3', and the ctRNA is complementary only to the 5' conserved sequence 5'-TTTTTGT-3'. This complementary sequence is located at a distance from the terminal loop of the ctRNA secondary structure. The ctRNA structure predicted by the mfold program suggests the possible generation of a terminal and an internal hairpin loop. The amount of in vitro translation product of repB mRNA was inversely proportional to the ctRNA concentration. Mutations in the terminal and internal hairpin loops of the ctRNA had inhibitory effects on its binding to the target mRNA. We propose that the intact structures of the terminal and internal hairpin loops, respectively, play important roles in forming the initial kissing and extending complexes between the ctRNA and target mRNA and that these regulate the copy number of this plasmid.
Keywords
ARBS; ctRNA; internal and terminal loops; operon; pJB01; RepB;
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Times Cited By KSCI : 1  (Citation Analysis)
Times Cited By Web Of Science : 6  (Related Records In Web of Science)
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