Title: | Identification of a conserved branched RNA structure that functions as a factor-independent terminator |
Author(s): | Johnson CM; Chen Y; Lee H; Ke A; Weaver KE; Dunny GM; |
Address: | "Department of Microbiology, University of Minnesota, Minneapolis, MN 55455" |
ISSN/ISBN: | 1091-6490 (Electronic) 0027-8424 (Print) 0027-8424 (Linking) |
Abstract: | "Anti-Q is a small RNA encoded on pCF10, an antibiotic resistance plasmid of Enterococcus faecalis, which negatively regulates conjugation of the plasmid. In this study we sought to understand how Anti-Q is generated relative to larger transcripts of the same operon. We found that Anti-Q folds into a branched structure that functions as a factor-independent terminator. In vitro and in vivo, termination is dependent on the integrity of this structure as well as the presence of a 3' polyuridine tract, but is not dependent on other downstream sequences. In vitro, terminated transcripts are released from RNA polymerase after synthesis. In vivo, a mutant with reduced termination efficiency demonstrated loss of tight control of conjugation function. A search of bacterial genomes revealed the presence of sequences that encode Anti-Q-like RNA structures. In vitro and in vivo experiments demonstrated that one of these functions as a terminator. This work reveals a previously unappreciated flexibility in the structure of factor-independent terminators and identifies a mechanism for generation of functional small RNAs; it should also inform annotation of bacterial sequence features, such as terminators, functional sRNAs, and operons" |
Keywords: | "Base Sequence Blotting, Northern Blotting, Western Conjugation, Genetic/genetics/*physiology Enterococcus faecalis/*genetics Molecular Sequence Data Mutagenesis Plasmids/*genetics RNA Folding/genetics RNA, Bacterial/*genetics Real-Time Polymerase Chain Re;" |
Notes: | "MedlineJohnson, Christopher M Chen, Yuqing Lee, Heejin Ke, Ailong Weaver, Keith E Dunny, Gary M eng R01 GM049530/GM/NIGMS NIH HHS/ GM49530/GM/NIGMS NIH HHS/ Research Support, N.I.H., Extramural 2014/02/20 Proc Natl Acad Sci U S A. 2014 Mar 4; 111(9):3573-8. doi: 10.1073/pnas.1315374111. Epub 2014 Feb 18" |