Summary
In Escherichia coli cells infected with bacteriophage ϕx174, mRNAs initiated by promoters PB and PD terminate after genes J, F, G, or H (TJ, TF, TG, or TH). These RNAs are relatively stable and contain mRNA-stabilizing signals at their 3′ ends. These signals were cloned after gene D of ϕx174 in an expression vector plasmid. The cloned signals stabilize mRNA of the upstream gene D and the stabilized mRNA is translationally functional. When these signals are inserted in reverse, no stabilizing effect on mRNA is observed indicating that the correct sequences at the 3′ ends of transcripts determine their stability. When a stabilizing signal (+) and a mutated stabilizing signal (-) which has reduced stabilizing activity are tandemly inserted after gene D, two sets of 3′ termini of the transcript are observed indicating that both signals also function as terminators. The amount of gpD synthesized from these constructs varies depending upon the relative positions of the (+) or (-) signals after gene D. The stabilizing function seems to act by preventing mRNA degradation from the 3′ to 5′ direction. Several common features of these stabilizers are described.
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References
Bechhofer DH, Dubnau D (1987) Induced mRNA stability in Bacillus subtilis. Proc Natl Acad Sci USA 84:498–502
Brennan SM, Geiduschek EP (1983) Regions specifying transcriptional termination and pausing in the bacteriophage SP01 terminal repeat. Nucleic Acids Res 11:4157–4175
Buckley KJ, Hayashi M (1987) Role of premature translational termination of expression of the ϕx174 lysis gene. J Mol Biol 198:549–607
Gorski K, Roch J-M, Prentki P, Krisch HM (1985) The stability of bacteriophage T4 gene 32 mRNA. A 5′ leader sequence that can stabilize mRNA transcripts. Cell 43:461–469
Guarneros G, Montanez C, Hernandez T, Court D (1982) Posttranscriptional control of bacteriophage lambda int gene expression from a site distal to the gene. Proc Natl Acad Sci USA 79:238–242
Hayashi M, Hayashi MN, Spiegelman S (1963) Restriction of in vivo genetic transcription to one of the complementary strands of DNA. Proc Natl Acad Sci USA 50:664–672
Hayashi M, Fujimura FK, Hayashi MN (1976) Mapping of in vivo messenger RNAs for bacteriophage ϕx174. Proc Natl Acad Sci USA 73:3519–3523
Hayashi MN, Hayashi M (1981) Stability of bacteriophage ϕx174 specific mRNA in vivo. J Virol 37:506–510
Hayashi MN, Hayashi M (1985) Cloned DNA sequences that determine mRNA stability of bacteriophage ϕx174 in vivo are functional. Nucleic Acids Res 13:5937–5948
Hayashi MN, Hayashi M, Müller UR (1983) Role for the J-F intercistronic region of bacteriophages ϕx174 and G4 in stability of mRNA. J Virol 48:186–196
Kennel DE (1986) In: Reznikoff WS, Gold WS (eds) Maximizing gene expression. Butterworth, Stoneham, Mass, pp 101–141
Kennel D, Canistraro VJ (1985) Evidence that the 5′ end of lac mRNA starts to decay as soon as it is synthesized. J Bacteriol 161:820–822
King TC, Sideskmuleh R, Schlessinger D (1986) Nucleolytic processing of ribonucleic acid transcripts in procaryotes. Microbiol Rev 50:428–451
Linney E, Hayashi M (1974) Intragenic regulation of the synthesis of ϕx174 gene A proteins. Nature 249:345–348
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Melefors Ö, von Gabain A (1988) Site-specific endonucleolytic cleavage and the regulation of stability of E. coli ompA mRNA. Cell 52:893–901
Mott JE, Galloway JL, Platt T (1985) Mutation of E. coli tryptophan operon mRNA: evidence for 3′ exonucleolytic processing after rho-dependent termination. EMBO J 4:1887–1891
Müller UR, Wells RD (1980a) Intercistronic regions in ϕx174 DNA. I. Construction of mutants with altered intercistronic regions between genes J and F. J Mol Biol 141:1–24
Müller UR, Wells RD (1980b) Intercistronic regions in ϕx174 DNA. II. Biochemical and biological analysis of mutants with altered intercistronic regions between genes J and F. J Mol Biol 141:25–41
Newbury SF, Smith NH, Robinson EC, Hiles ID, Higgins CF (1987a) Stabilization of translationally active mRNA by prokaryotic REP sequences. Cell 48:247–310
Newbury SF, Smith NH, Higgins CF (1987b) Differential mRNA stability controls relative gene expression within a polycistronic operon. Cell 51:1131–1143
Sanger F, Coulson AR, Friedman T, Air GM, Barrell BC, Brown NL, Fiddes JC, Hutchison III CA, Slocombe PM, Smith M (1978) The nucleotide sequence of bacteriophage ϕx174. J Mol Biol 125:225–246
Schindler D, Echols H (1981) Retroregulation of the int gene of bacteriophage lambda: Control of translation completion. Proc Natl Acad Sci USA 78:4475–4479
Schmeissner V, McKenney K, Rosenberg M, Court D (1984) Removal of terminator structure by RNA processing regulates in gene expression. J Mol Biol 176:39–52
Wong HC, Chang S (1986) Identification of a positive retroregulator that stabilizes mRNA in bacteria. Proc Natl Acad Sci USA 83:3233–3237
Zuker M, Stiegler P (1981) Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Res 9:133–148
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Communicated by J.W. Lengeler
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Hayashi, M.N., Yaghmai, R., McConnell, M. et al. mRNA stabilizing signals encoded in the genome of the bacteriophage ϕx174. Mol Gen Genet 216, 364–371 (1989). https://doi.org/10.1007/BF00334377
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DOI: https://doi.org/10.1007/BF00334377