Function of the novel subdomain in the RNA binding domain of transcription termination factor Rho from Micrococcus luteus.
Transcription termination factor Rho from Micrococcus luteus, a high G + C Gram-positive bacterium, contains an unusual extra sequence within its RNA binding domain that is rich in Arg, Glu, and Asp residues and deficient in hydrophobic residues. To determine the role of this extra sequence, we compared the biochemical properties of a variant lacking nearly all the extra sequence, des(60-300) Rho, to that of wild-type M. luteus Rho. The two forms had very similar properties except that the des(60-300) Rho was unable to terminate transcription with Escherichia coli RNA polymerase at the promoter proximal sites used by the wild-type Rho on a lambda cro DNA template but could cause termination at more distal sites and did cause termination at proximal sites when ITP replaced GTP in the reaction mixture. The RNA binding properties of the two forms of this Rho with normal and inosine-substituted RNAs were found to correlate fully with their termination properties. These results indicate that the arginine-rich extra sequence is directly involved in the selection of the termination site and support the hypothesis that the sequence is present in M. luteus Rho to facilitate its binding to M. luteus transcripts, which are likely to have a high degree of base-paired secondary structure because of their high proportion of G residues.
Nowatzke
W L
WL
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.
Burns
C M
CM
Richardson
J P
JP
D000251:Adenosine Triphosphatases / Q000378:metabolism; D001665:Binding Sites; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006160:Guanosine Triphosphate / Q000378:metabolism; D007293:Inosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D016982:Micrococcus luteus; D009690:Nucleic Acid Conformation; D012313:RNA / Q000378:metabolism*; D012234:Rho Factor / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic
D012234:Rho Factor; D007293:Inosine Triphosphate; D012313:RNA; D006160:Guanosine Triphosphate; D000251:Adenosine Triphosphatases; C031715:rho-ATPase