Requirement of chain initiation factor 3 and ribosomal protein S1 in translation of synthetic and natural messenger RNA

Sobura, Joseph; Chowdhury, Madhumita Roy; Hawley, David; Wahba, Amy

doi:10.1093/nar/4.1.17

Cited by 44 publications

(15 citation statements)

References 36 publications

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“…these results do not rule out IF3 (6,8) or ribosomal S1 (8,29) interaction with mRNA, since both of these proteins may have two RNA binding sites (21,48). Hence, it will be of interest to look for any similarly strong interactions of IF3 with the same or other regions of 16S RNA in whole 30S ribosomal subunits, where the 3' or 5' termini have been radiolabeled in aitil.…”

Section: G~~~~~-mentioning

confidence: 85%

Nuclease mapping of the secondary structure of the 49-nucleotide 3' terminal cloacin fragment ofEscherichia coli16S RNA and its interactions with initiation factor 3

Wickstrom¹

1983

Nucl Acids Res

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Escherichia coli translational initiation factor 3 (IF3) may be crosslinked to the 3' end of 16S RNA in 30S ribosomal subunits. In order to determine the sequence to which IF3 may bind in vivo, samples of 5'-32P labelled 3' terminal 49-nucleotide fragment of 16S RNA were incubated 5 min. at 37 degrees in 40 mM Tris-HOAc, pH 7.4, 100 mM NaCl, 1 mM Mg (OAc)2, 1 mM ZnSO4, with or without IF3, then reacted a further 5 min with nuclease S1, RNase T1, or RNase A. Base pairing between the 5' and 3' legs of the fragment occurs in the absence of IF3, but is disrupted by IF3 binding. IF3 appears to protect some residues near the 5' end of the fragment (U1495, A1499, A1500, A1502, and A1503) from nuclease S1, and potentiates S1 attack on others (G1494, G1497, C1501, G1504, G1505, U1506, G1517, G1529, G1530, and C1533). A series of equimolar reactions at increasing dilution imply an association constant range of 1.4-7.0 X 10(7) M-1.

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Section: G~~~~~-mentioning

confidence: 85%

Nuclease mapping of the secondary structure of the 49-nucleotide 3' terminal cloacin fragment ofEscherichia coli16S RNA and its interactions with initiation factor 3

Wickstrom¹

1983

Nucl Acids Res

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“…The fingerprints of Figure 1 contain not only spots derived from the three authentic initiator regions in the R17 RNA (see Table 1) but other large oligonucleotides as well (numbers 10,11,12,13). These are prominent only in the ribosome-protected RNA from reactions where Sl was not present (Figure lc and d); they are barely detectable in maps from reactions containing either 30S(-Sl) subunits reconstituted with S1 (Figure la and b) or native 30S subunits (not shown).…”

Section: Sl Decreases Ribosome Binding To Spurious Sites In R17 Rnamentioning

confidence: 99%

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

et al. 1977

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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.

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“…to 70S ribosomes was demonstrated by analyzing ['4C] IF3-70S mixtures on polyacrylamide gel electrophoresis (Sobura and Wahba, unpublished observations). Furthermore, the fact that IF3 is required for maximum translation at 12-14 mM Mg2+ (Sobura et al, 1977) strongly suggests an interaction between IF3 and 70S ribosomes.…”

Section: Resultsmentioning

confidence: 99%

Kinetic studies of the rates and mechanism of assembly of the protein synthesis initiation complex

Goss¹,

Parkhurst²,

Wahba³

1980

Biophysical Journal

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Rate constants for a number of the assembly reactions involved in forming Escherichia coli ribosome initiation complexes have been measured. These reactions were monitored in a stopped-flow device in which Rayleigh scattering and fluorescence anisotropy were followed as a function of time. Fluorescence was induced by laser excitation modulated at 50 kHz. Aminoacyl-tRNA, initiation factor 3 (IF3), and 70S ribosomes were labeled with fluorescent probes. The light-scattering and fluorescence data show that the antiassociation model for IF3 function cannot be correct. IF3 can be considered to act as an effector in an allosteric model for ribosome function. Fluorescence anisotropy stopped-flow experiments provided rate constants for the binding of IF3 to both 30S subunits and to the intact 70S ribosome. Aminoacyl-tRNA's and nucleotide triplets appear to bind rapidly to 70S ribosomes and then a slow first-order conformational change occurs.

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Requirement of chain initiation factor 3 and ribosomal protein S1 in translation of synthetic and natural messenger RNA

Cited by 44 publications

References 36 publications

Nuclease mapping of the secondary structure of the 49-nucleotide 3' terminal cloacin fragment ofEscherichia coli16S RNA and its interactions with initiation factor 3

Nuclease mapping of the secondary structure of the 49-nucleotide 3' terminal cloacin fragment ofEscherichia coli16S RNA and its interactions with initiation factor 3

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

Kinetic studies of the rates and mechanism of assembly of the protein synthesis initiation complex

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