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

Steitz, Joan A.; Wahba, Amy; Laughrea, Michael; Moore, Peter B.

doi:10.1093/nar/4.1.1

Cited by 76 publications

(24 citation statements)

References 56 publications

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“…We have observed that, in the absence of ribosomal salt wash fraction, the synthesis of specific proteins encoded by mRNA derived from the bacillus phage 429 is 40% of that in the presence of factors present in a salt wash of ribosomes in translation systems derived from both E. coli and B. subtilis. This contrasts sharply with the complete dependence on the salt wash for translation by E. coli ribosomes of T7 and other mRNAs derived from Gram-negative organisms (10,(36)(37)(38).…”

Section: Discussionmentioning

confidence: 85%

“…S1 than is recognition of the A protein site which has a complementarity of.seven base-pairs (37). 'It has been suggested that the high degree of complementarity between the A site and the 3' end of 16S rRNA may provide sufficient binding energy.…”

Section: Discussionmentioning

confidence: 99%

“…'It has been suggested that the high degree of complementarity between the A site and the 3' end of 16S rRNA may provide sufficient binding energy. to obviate the usual factor requirements (37 …”

Section: Discussionmentioning

confidence: 99%

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Initiation factor-independent translation of mRNAs from Gram-positive bacteria.

McLaughlin¹,

Murray²,

Rabinowitz³

1981

Proc. Natl. Acad. Sci. U.S.A.

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Initiation factor-independent translation of mRNA derived from bacillus phage 429 DNA occurs with translation systems derived from Bacilus subtilis or Escherichia coli. This is in sharp contrast to the strict dependence on ribosome salt wash fraction ofE. coli ribosomes for the translation ofT7 and other mRNAs derived from Gram-negative organisms.Although the classification of bacteria as Gram-negative or Gram-positive is based on differences in cell wall architecture (1), additional distinctions between these two groups ofbacteria are known. Evolutionary trees based on 16S and 5S RNA sequences (2, 3) indicate that the Gram-positive bacteria represent some of the most primitive species, such as the clostridia, that are characterized by fermentative metabolism and the ability to form endospores in a hostile environment. The Grampositive bacilli appeared somewhat later with perhaps the first use of aerobic respiratory metabolism, but they retain sporeforming ability. The Gram-negative bacteria evolved even later. They are often capable offacultative respiratory metabolism but have lost the ability to sporulate. Instead, the Gram-negative bacteria, such as Escherichia coli, are equipped to adapt to a changing environment.One manifestation of the increased adaptability of E. coli is its ability to express genes from a wide variety of bacteria including both Gram-positive and Gram-negative species. In contrast, many E. coli genes carried on hybrid plasmids are not expressed in the Gram-positive species Bacillus subtilis (4-6). These in vivo restrictions of heterospecific gene expression in B. subtilis could result from limitations in transcription, translation, or posttranslational processing of genetic information.There is increasing evidence that the translational machinery isolated from E. coli differs significantly from that of its Grampositive predecessor, B. subtilis. Studies from several laboratories indicate that ribosomes isolated from Gram-positive bacteria are extremely inefficient in translating mRNAs derived from Gram-negative bacteria (7-13). Ribosomes from E. coli, on the other hand, show comparable translational efficiency in response to mRNAs from Gram-positive or Gram-negative bacteria.Reconstitution ofhybrid 30S ribosomal subunits has revealed differences in the protein components which affect the efficiencies with which Gram-negative mRNAs are translated (11,14,15). For example, Isono and Isono (16) reported that translation of f2 RNA by Bacillus stearothermophilus ribosomes is stimulated by the addition of E. coli ribosomal protein S1. We have demonstrated here that E. coli ribosomal protein S1 has very little effect on the efficiency with which f2 RNA and T7 mRNA are translated by the B. subtilis translation system and concluded that S1 is not the sole determinant ofspecies-specific initiator recognition.Another approach to understanding the basis for selective mRNA recognition by B. subtilis ribosomes involves characterization of their interaction with mRNAs from Gram-positive organism...

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Section: Discussionmentioning

confidence: 85%

Section: Discussionmentioning

confidence: 99%

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Initiation factor-independent translation of mRNAs from Gram-positive bacteria.

McLaughlin¹,

Murray²,

Rabinowitz³

1981

Proc. Natl. Acad. Sci. U.S.A.

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“…In prokaryotes, substantial evidence indicates that a region located about 10 nucleotides 5' to the initiation codon is involved in base pairing with the 3'-terminal sequence of the 16S rRNAs (29)(30)(31). Similar base pairing in eukaryotes between the 3' terminus of the 18S rRNA and the untranslated region of the mRNA has also been suggested (32)(33)(34).…”

Section: Resultsmentioning

confidence: 91%

The nucleotide sequences of the untranslated 5' regions of human alpha- and beta-globin mRNAs.

Chang¹,

Temple²,

Poon³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

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The complete sequences of the untranslated 5' regions of human a-and ftlobin mRNAs were determined by sequence analysis of full-length cDNAs. The single-stranded cDNAs were digested with the restriction endonuclease Hae Ill, and the two 3'-terminal fragments of 75 (5)(6)(7)(8)(9)(10)(11)(12). In the human, the ,Bglobin mRNA from the initiation codon AUG to the 3'-poly(A) end has been sequenced, and portions of the translated region of the a-globin mRNA and the complete untranslated 3' end are known (10, 13-16). The primary structure of the untranslated 5' region of mRNA is of great interest because it may play a role in ribosomal binding and the initiation of protein synthesis. Lockard and RajBhandary (17) determined the sequence of the 5' termini of the separated rabbit a-and ,B-globin mRNAs by direct RNA sequence analysis. By analyzing the cDNAs synthesized with a primer complementary to the region of the initiation codon, Baralle (11,12) sequenced the complete untranslated 5' regions of both rabbit a-and ,B-globin mRNAs.We report here the complete sequences of the untranslated 5' region of the human a-and,B-globin mRNAs. We utilized the ability of the restriction enzyme Hae III to cleave singlestranded DNA at G-G-C-C sequences (18,19). Full-length single-stranded a-and ,B-globin cDNAs were digested with Hae III and the two 3'-terminal fragments corresponding to the 5' termini of the a-and f3-mRNAs were identified and isolated.The sequences of these two fragments were then determined according to the method of Maxam and Gilbert (4) by labeling either at their 3' ends with ribonucleoside [32P]diphosphate with terminal deoxyribonucleotidyl transferase, or at their 5' ends with 32P by using polynucleotide kinase.MATERIALS AND METHODS Source of Human Globin mRNA. Blood was obtained from a patient with sickle cell disease, one with hemoglobin H disease, and one with homozygous 3 thalassemia. RNA was prepared from these samples and the poly(A)-rich RNA was isolated by two passages over oligo(dT)-cellulose as described (20).Preparation of Globin cDNAs. Single-stranded globin cDNAs were prepared according to the method of Verma et al. (21). Nonradioactive dATP, dGTP, and TTP at 400,M each and radioactive dCTP at 100 MM were used to maximize the synthesis of full-length cDNAs as shown by Efstratiadis et al. (22). The radioactive precursors used for the synthesis of internally labeled cDNA in the different experiments were [3H]dCTP (New England Nuclear, 23 Ci/mmol) diluted to a specific activity of 2.3 Ci/mmol and [a-32P]dCTP (New England Nuclear, 110-115 Ci/mmol) diluted to a specific activity of 25 or 0.1 Ci/mmol to yield high-or low-specific activity 32p products, respectively.Digestion of cDNA with Restriction Enzyme. cDNA was digested with the restriction endonuclease Hae III (New England Biolabs), 200 units/Mg of DNA, in 6 mM Tris-HCI, pH 7.4/6 mM MgCl2/6 mM NaCl/6 mM 2-mercaptoethanol for 16 hr at 37°. To analyze the restriction fragments, digested high-specific activity [32P]cDNA was separated by ...

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“…It seems clear, however, that the 16 S RNA does play a role in IF-3 binding [ 1 ] . Furthermore, IF-3 and the 3'-end of the 16 S RNA appear to be functionally [4] as well as topographically related [S] . These results, as well as speculations about the cause of the IF-~'S dissociation activity [6] , make plausible the idea that some of the nucleotides adjacent to the 3'-end of 16 S RNA may be in the immediate nei~borhood of IF-3, or that they may be an important part of the IF-3 binding site on the 30 S subunit.…”

Section: Introductionmentioning

confidence: 99%