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M Laughrea

Publications and source records attributed to M Laughrea.

38 records · Page 3Linked to original sources

Differential requirements for polypeptide chain initiation complex formation at the three bacteriophage R17 initiator regions.

The initiation specificity of washed E. coli ribosomes in the presence and absence of purified initiation factors and/or S1 protein has been examined in protection experiments using 32P-labelled R17 RNA. We find that the three bacteriophage initiator regions do not exhibit equal requirements for either of these components during initiation complex formation. Specifically, both factors and S1 stimulate ribosome binding to the beginnings of the coat and replicase cistrons to a greater extent than they promote recognition of the A protein initiation site. The differential effects are therefore inversely correlated with the degree of mRNA-16S rRNA complementarity exhibited by the three initiator regions. We also observe that S1 suppresses ribosome binding to spurious sites in the R17 RNA.

Bacterial Proteins↗

Ribosomal protein S1 and initiation factor IF3 do not promote the ribosomal binding of approximately 19-nucleotide-long mDNA and mRNA models.

A number of model mDNAs and mRNAs, about 19 nucleotides in length, were obtained by automated synthesis and T7 RNA polymerase directed transcription from synthetic DNA templates, respectively. These mDNAs and mRNAs had Shine-Dalgarno sequences that were four to eight nucleotides long and their sequence was similar to the R17 coat protein initiation site. The effect of S1 or initiation factors (IFs) on the rate and extent of ribosomal binding of these mDNAs and mRNAs was investigated by nitrocellulose filtration. No effect was detected, suggesting that the main function of S1 or IFs included neither the direct recognition or binding of short messengers to the 30S subunits, nor their rejection as "false" for possessing the wrong sugars or an insufficient length. A pulse-chase experiment with one of the mRNAs also showed that S1 or IF3 did not influence the exchange rate of mRNA bound to the 30S subunit.

Bacterial Proteins↗