title: Precursor of C4 antisense RNA of bacteriophages P1 and P7 is a substrate for RNase P of Escherichia coli.
abstract: The C4 repressor of the temperate bacteriophages P1 and P7 inhibits antirepressor (Ant) synthesis and is essential for establishment and maintenance of lysogeny. C4 is an antisense RNA acting on a target, Ant mRNA, which is transcribed from the same promoter. The antisense-target RNA interaction requires processing of C4 RNA from a precursor RNA. Here we show that 5' maturation of C4 RNA in vivo depends on RNase P. In vitro, Escherichia coli RNase P and its catalytic RNA subunit (M1 RNA) can generate the mature 5' end of C4 RNA from P1 by a single endonucleolytic cut, whereas RNase P from the E. coli rnpA49 mutant, carrying a missense mutation in the RNase P protein subunit, is defective in the 5' maturation of C4 RNA. Primer extension analysis of RNA transcribed in vivo from a plasmid carrying the P1 c4 gene revealed that 5'-mature C4 RNA was the predominant species in rnpA+ bacteria, whereas virtually no mature C4 RNA was found in the temperature-sensitive rnpA49 strain at the restrictive temperature. Instead, C4 RNA molecules carrying up to five extra nucleotides beyond the 5' end accumulated. The same phenotype was observed in rnpA+ bacteria which harbored a plasmid carrying a P7 c4 mutant gene with a single C-->G base substitution in the structural homologue to the CCA 3' end of tRNAs. Implications of C4 RNA processing for the lysis/lysogeny decision process of bacteriophages P1 and P7 are discussed.
authors.affiliation: Abteilung Erdmann, Freie Universität Berlin, Germany.
authors.forename: R K
authors.initials: RK
authors.lastname: Hartmann
authors.forename: J
authors.initials: J
authors.lastname: Heinrich
authors.forename: J
authors.initials: J
authors.lastname: Schlegl
authors.forename: H
authors.initials: H
authors.lastname: Schuster
mesh_terms: D017101:Bacteriophage P1 / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003090:Coliphages / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D004722:Endoribonucleases / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D012322:RNA Precursors / Q000378:metabolism*; D016372:RNA, Antisense / Q000096:biosynthesis* / Q000737:chemistry; D016337:RNA, Catalytic / Q000302:isolation & purification / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D012353:RNA, Transfer, Gly / Q000096:biosynthesis / Q000302:isolation & purification; D012367:RNA, Viral / Q000096:biosynthesis* / Q000737:chemistry; D012097:Repressor Proteins / Q000096:biosynthesis*; D043262:Ribonuclease P; D014158:Transcription, Genetic
chemical_list: D017931:DNA Primers; D029968:Escherichia coli Proteins; D012322:RNA Precursors; D016372:RNA, Antisense; D016337:RNA, Catalytic; D012333:RNA, Messenger; D012353:RNA, Transfer, Gly; D012367:RNA, Viral; D012097:Repressor Proteins; D004722:Endoribonucleases; D043262:Ribonuclease P; C470865:ribonuclease P, E coli