TY - JOUR
T1 - Autogenous regulatory site on the bacteriophage T4 gene 32 messenger RNA
AU - McPheeters, David S.
AU - Stormo, Gary D.
AU - Gold, Larry
N1 - Funding Information:
We thank Roger Saunders and Tom Schneider for construction of the plasmids pRS210 and pTS26. We also thank Roger Saunders for the secondary structure analysis of the mRNA encoded by pRS210. We also thank Tom Cech for generously providing the densitometer for our use. We appreciate the helpful conversations with Olke Uhlenbeck, Ignacio Tinoco, and John Milligan about pseudoknots. A timely letter from Dr C. W. A. Pleij led us to understand pseudoknots more fully, and to generate Figure 14. Drew Smith helped with the molecular graphics of Figure 14(b), using the program MIDAS from Robert Langridge at UCSF. Finally, Craig Tuerk, Mike Yarus, Nancy Guild, Kathy Krassa and Peter Gauss provided helpful suggestions. This work was supported by NIH grants GM28685 and GM19963 to L.G.
PY - 1988/6/5
Y1 - 1988/6/5
N2 - We have identified the binding site on the bacteriophage T4 gene 32 mRNA responsible for autogenous translational regulation. We demonstrate that this site is largely unstructured and overlaps the initiation codon of gene 32 as previously predicted. Co-operative binding of gene 32 protein to this site specifically blocks the formation of 30 S-tRNAmetf-gene 32 mRNA ternary complexes and initiation of translation. The translational operator is bound co-operatively by gene 32 protein and this binding is facilitated by a nucleation site far upstream from the initiation codon. A similar unstructured mRNA lacking this nucleation site is also bound co-operatively, but only at concentrations of gene 32 protein higher than those needed to repress binding of ribosomes to the gene 32 mRNA. Some sequence-specific interactions may also influence this binding. Comparison of the bacteriophage T2, T4 and T6 gene 32 operator sequences leads us to propose that the nucleation site is a pseudoknot.
AB - We have identified the binding site on the bacteriophage T4 gene 32 mRNA responsible for autogenous translational regulation. We demonstrate that this site is largely unstructured and overlaps the initiation codon of gene 32 as previously predicted. Co-operative binding of gene 32 protein to this site specifically blocks the formation of 30 S-tRNAmetf-gene 32 mRNA ternary complexes and initiation of translation. The translational operator is bound co-operatively by gene 32 protein and this binding is facilitated by a nucleation site far upstream from the initiation codon. A similar unstructured mRNA lacking this nucleation site is also bound co-operatively, but only at concentrations of gene 32 protein higher than those needed to repress binding of ribosomes to the gene 32 mRNA. Some sequence-specific interactions may also influence this binding. Comparison of the bacteriophage T2, T4 and T6 gene 32 operator sequences leads us to propose that the nucleation site is a pseudoknot.
UR - http://www.scopus.com/inward/record.url?scp=0024278647&partnerID=8YFLogxK
U2 - 10.1016/0022-2836(88)90634-1
DO - 10.1016/0022-2836(88)90634-1
M3 - Article
C2 - 3262167
AN - SCOPUS:0024278647
SN - 0022-2836
VL - 201
SP - 517
EP - 535
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 3
ER -