The sequences and activities of RegB endoribonucleases of T4-related bacteriophages.
The RegB endoribonuclease encoded by bacteriophage T4 is a unique sequence-specific nuclease that cleaves in the middle of GGAG or, in a few cases, GGAU tetranucleotides, preferentially those found in the Shine-Dalgarno regions of early phage mRNAs. In this study, we examined the primary structures and functional properties of RegB ribonucleases encoded by T4-related bacteriophages. We show that all but one of 36 phages tested harbor the regB gene homologues and the similar signals for transcriptional and post-transcriptional autogenous regulation of regB expression. Phage RB49 in addition to gpRegB utilizes Escherichia coli endoribonuclease E for the degradation of its transcripts for gene regB. The deduced primary structure of RegB proteins of 32 phages studied is almost identical to that of T4, while the sequences of RegB encoded by phages RB69, TuIa and RB49 show substantial divergence from their T4 counterpart. Functional studies using plasmid-phage systems indicate that RegB nucleases of phages T4, RB69, TuIa and RB49 exhibit different activity towards GGAG and GGAU motifs in the specific locations. We expect that the availability of the different phylogenetic variants of RegB may help to localize the amino acid determinants that contribute to the specificity and cleavage efficiency of this processing enzyme.
Piesiniene
Lina
L
Department of Gene Engineering, Institute of Biochemistry, Mokslininku 12, 08662 Vilnius, Lithuania.
Truncaite
Lidija
L
Zajanckauskaite
Aurelija
A
Nivinskas
Rimas
R
D000595:Amino Acid Sequence; D017122:Bacteriophage T4 / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D004722:Endoribonucleases / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity; D014158:Transcription, Genetic / Q000235:genetics
D012333:RNA, Messenger; D012367:RNA, Viral; D004722:Endoribonucleases; C407103:T4 RegB endoribonuclease