Summary
In protein synthesis, the incorporation of an N-terminal formylmethionine residue is directed by an initiation codon. The most frequently used codon is AUG, although initiation at GUG and UUG codons has also been observed. The HD263 mutation is an AUG to AUA change in the rIIB initiation codon. Evidence is presented here that wild type and HD263 rIIB proteins, whether synthesized in vivo or in vitro, have identical fmet peptides. It is concluded that translation began at the AUA mutant initiation codon in vitro and in phage T4 infected cells.
In the in vitro translation system used in these studies, the rIIB protein synthesized at 25° no longer contains the N-terminal formyl group whereas a large proportion of the formyl group is retained at 37°.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- tss-mutation:
-
temperature-sensitive, synthesis mutation
- PrIIB:
-
protein product of gene rIIB
- PrIIB+ :
-
PHD263 and PHE122, rIIB proteins synthesized by rIIB+ phage, tss-mutant HD263 and amber mutant HE122
- fmet-tRNA:
-
N-formylmethionyl-tRNA metinf
References
Adams, J.M.: On the release of the formyl group from nascent protein. J. Mol. Biol. 33, 571–589 (1968)
Adams, J.M., Capecchi, M.R.: N-formyl-methionyl-sRNA as the initiator of protein synthesis. Proc. Natl. Acad. Sci. U.S.A. 55, 147–155 (1966)
Allet, B., Katagiri, K.J., Gesteland, R.F.: Characterization of polypeptides made in vitro from bacteriophage Lambda DNA. J. Mol. Biol. 78, 589–600 (1973)
Ball, L.A., Kaesberg, P.: Cleavage of the N-terminal formylmethionine residue from a bacteriophage coat protein in vitro. J. Mol. Biol. 79, 531–537 (1973)
Barnett, L., Brenner, S., Crick, F., Shulman, R., Watts-Tobin, R.: Phase-shift and other mutants in the first part of the rIIB cistron of bacteriophage T4. Philos. Trans. R. Soc. 252, 487–560 (1967)
Belin, D., Epstein, R.H.: A temperature-sensitive rIIB mutation which affects the synthesis of bacteriophage T4 rIIB protein. Virology 78, 537–553 (1977)
Belin, D., Hedgpeth, J., Selzer, G., Epstein, R.H.: Temperature-sensitive mutation in the initiation codon of the rIIB gene of bacteriophage T4. Proc. Natl. Acad. Sci. U.S.A., in press (1979)
Brenner, S., Beckwith, J.R.: Ochre mutants, a new class of suppressible nonsense mutants. J. Mol. Biol. 13, 629–637 (1965)
Capecchi, M.R.: Initiation of E. coli proteins. Proc. Natl. Acad. Sci. U.S.A. 55, 1517–1524 (1966)
Clark, B.F.C., Marcker, K.A.: The role of N-formyl-methionyl-sRNA in protein biosynthesis. J. Mol. Biol. 17, 394–406 (1966)
Champe, S.P., Benzer, S.: Reversal of mutant phenotypes by 5-fluorouracil: an approach to nucleotide sequences in messenger RNA. Proc. Natl. Acad. Sci. U.S.A. 48, 532–546 (1962)
Dunn, J.J., Buzash-Pollert, E., Studier, F.W.: Mutations of bacteriophage T7 that affect initiation of synthesis of the gene 0.3 protein. Proc. Natl. Acad. Sci. U.S.A. 75, 2741–2745 (1978)
Edgar, R.S. and Epstein, R.H.: Conditional lethal mutations in bacteriophage T4. In: Genetics today (S.J. Geerts, ed.), pp. 1–17. Oxford: Pergamon Press 1964
Epstein, R.H., Bolle, A., Steinberg, C.M., Kellenberger, E., Boy de la Tour, E., Chevalley, R., Edgar, R.S., Susman, M., Denhardt, G.H., Lielausis, A.: Physiological studies of conditional lethal mutants of bacteriophage T4D. Cold Spring Harbor Symp. Quant. Biol. 28, 375–394 (1963)
Files, J.G., Weber, K., Coulondre, C., Miller, J.H.: Identification of the UUG codon as a translational initiation codon in vivo. J. Mol. Biol. 95, 327–330 (1975)
Gesteland, R.F.: Isolation and characterization of ribonuclease I mutants of Escherichia coli. J. Mol. Biol. 16, 67–84 (1966)
Katz, E.R., Brenner, S.: Genetic map of the beginning of the rIIB cistron of bacteriophage T4. Mol. Gen. Genet. 143, 101–104 (1975)
Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970)
Livingston, D.M., Leder, P.: Deformylation and protein synthesis. Biochemistry 8, 435–443 (1969)
Marcker, K.: The formation of N-formyl-methionyl-sRNA. J. Mol. Biol. 14, 63–70 (1965)
Singer, B.S., Gold, L.: A mutation that confers temperature sensitivity on the translation of rIIB in bacteriophage T4. J. Mol. Biol. 103, 627–646 (1976)
Steege, D.A.: 5-terminal nucleotide sequence of E. coli lactose repressor mRNA: features of translational initiation and reinitiation sites. Proc. Natl. Acad. Sci. U.S.A. 74, 4163–4167 (1977)
Steitz, J.A.: Genetic signals and nucleotide sequences in messenger RNA. In: Biological regulation and control, Vol. 1 (R. Goldberger, ed.), New York: Plenum Publishing Co. in press 1978
Webster, R.E., Engelhardt, D.L., Zinder, N.: In vitro protein synthesis: Chain initiation. Proc. Natl. Acad. Sci. U.S.A. 55, 155–161 (1966)
Zubay, G., Chambers, D.A., Cheong, L.C.: Cell-free studies on the regulation of the lac operon. In: The lactose operon (J.R. Beckwith and D. Zipser, eds.), pp 375–391. Cold Spring Harbor (New York): Cold Spring Harbor Laboratory 1970
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
Rights and permissions
About this article
Cite this article
Belin, D. Bacteriophage T4 rIIB protein synthesis with a temperature-sensitive mutation in the rIIB initiation codon. Molec. Gen. Genet. 171, 35–42 (1979). https://doi.org/10.1007/BF00274012
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00274012