Redundancy of primary RNA-binding functions of the bacterial transcription terminator Rho.
Bottom Line: The bacterial transcription terminator, Rho, terminates transcription at half of the operons.These terminators function optimally only through a NusG-assisted recruitment and activation of Rho.Our data also indicate that at these terminators, Rho-EC-bound NusG interaction facilitates the isomerization of Rho into a translocase-competent form by stabilizing the interactions of mRNA with the secondary RNA binding site, thereby overcoming the defects of the primary RNA binding functions.
Affiliation: Laboratory of Transcription, Center for DNA Fingerprinting and Diagnostics, Tuljaguda Complex, 4-1-714 Mozamjahi Road, Nampally, Hyderabad 500001, India.Show MeSH
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Mentions: To study the differential growth behavior between the WT and different rho mutants (Figure 1), MG1655rac+strains (having the rac prophage) were at first transformed with WT and different Rho mutants expressed from a low copy number plasmid, pCL1920 (pHYD567), and, subsequently, the chromosomal rho was removed by inserting a kanR cassette (rho::kanR) via P1 transduction (Supplementary Figure S4A). The transductants were re-streaked on the LB plates supplemented with appropriate antibiotics (Figure 1B). In this rho::kanR cassette, first 360 bases of the rho sequence are replaced with the kanamycin resistance gene, and hence rho is linked very tightly with this marker (P1 was made on the strain RS330; see Table 1). We have also confirmed the absence of duplication of rho in all the transductants by PCR using rho specific primers (Supplementary Figure S4B).
Affiliation: Laboratory of Transcription, Center for DNA Fingerprinting and Diagnostics, Tuljaguda Complex, 4-1-714 Mozamjahi Road, Nampally, Hyderabad 500001, India.