Ribosomal proteins of bacterial cells: Strain- and species-specificity

Otaka, Eiko; Itoh, Takuzì; Osawa, Syozo

doi:10.1016/0022-2836(68)90283-0

Cited by 92 publications

(25 citation statements)

References 14 publications

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“…le) (12). Unresolved protein components were fractionated on carboxymethylcellulose at pH 5.6 (13). Proteins were identified by polyacrylamide gel electrophoresis; peak fractions were pooled, concentrated by ultrafiltration, and stored at -20°.…”

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confidence: 99%

Location of Ribosomal Protein Binding Sites on 16S Ribosomal RNA

Zimmermann

Muto

Fellner

et al. 1972

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

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The distribution of ribosomal protein binding sites on the 16S ribosomal RNA molecule has been analyzed by limited ribonuclease hydrolysis of RNA-protein complexes, as well as by the interaction of individual proteins with RNA fragments purified from partial enzymatic digests. Of the six 30S subunit proteins known to interact directly with 16S RNA, proteins S4, S8, S15, S20, and, probably, S13 bind within a fragment produced by T1 RNase (12S RNA) that comprises some 900 nucleotides and covers almost the entire 5'-terminal half of the 16S molecule. A fragment of 500-00 nucleotides (8S RNA) that is contiguous with 12S RNA and arises from the 3'-terminal portion of the 16S molecule is believed to contain the binding site for protein S7. Protein S15 interacts specifically with a sequence of about 135 nucleotides (4S RNA) that derives from 12S RNA after more extensive hydrolysis. Protein S4, but none of the other ribosomal proteins, binds to a 500-nucleotide fragment (9S RNA), generated by pancreatic RNase, that lies at the 5'-terminus of 16S RNA and is completely overlapped by the 12S fragment. A preliminary map of the binding sites is presented.Reconstitution of the 30S ribosomal subunit of Escherichia coli proceeds by the sequential and cooperative addition of 20 or 21 different proteins to 16S RNA (1). In the initial stages of the assembly sequence, six ribosomal proteins directly interact with independent binding sites on the RNA molecule (1-4). The specific structural features that permit ribosomal RNA to recognize and bind these proteins are not known.We ratio is increased to 1:5, hydrolysis is more extensive; fragments of about 10 S and 4 S are generated from the 12S RNA, as demonstrated by nucleotide sequence analysis, and the 8S RNA is degraded to small oligonucleotides. Fragments with the same sedimentation rates result when enzymatic digestion is performed after individual ribosomal proteins have been attached to the 16S RNA (Fig. 1, dashed curves).The action of pancreatic ribonuclease A on 16S RNA is also limited under similar conditions, and the presence of bound protein again affords the RNA no additional protection. At an enzyme to substrate ratio of 1:20, the principal component in the digest sediments at about 11 S; an increase of the ratio to 1: 5 results in fragments with apparent sedimentation coefficients of 9 S and 4 S.The resistance of large RNA segments to RNase digestion in these experiments is attributable to the high concentration of Mg++ ions in the incubation buffer; reduction of the Mg++ concentration alone renders the 16S RNA more labile to degradation, whereas removal of mofnovalent cations has no effect on fragment size. Since the RNA chain possesses a high degree of secondary structure and a compact conformation at the Mg++ concentrations used (6), RNase attack is most likely limited by decreased accessibility of the 16S molecule to the enzyme. (Fig. lb), while no protein S4 radioactivity is observed in the 8S peak. Under the same conditions, S8, S13, S15, and S20, four oth...

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mentioning

confidence: 99%

Location of Ribosomal Protein Binding Sites on 16S Ribosomal RNA

Zimmermann

Muto

Fellner

et al. 1972

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

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“…When the elution patterns of [l4C]lysine-labeled ribosomal proteins of Escherichia coli Q13 were compared with those of [3H]lysine-Jabeled ribosomal proteins of E coli K12, they proved to be identical (41). No apparent similarities could be detected in comparisons of E coli ribosomal proteins with those of several Bacillus species and with Sarcina.…”

Section: Ribosomal Protein Patterns and Genetic Relatednessmentioning

confidence: 92%

Phototrophic Green and Purple Bacteria: A Comparative, Systematic Survey

Pfennig¹

1977

Annu. Rev. Microbiol.

144

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“…Several researchers (19,26,27) have suggested that acrylamide gel electrophoresis of ribosomal proteins might be a useful taxonomic tool. The results here suggest that disc gel electrophoresis data of 50s ribosomal proteins alone are not adequate taxonomic criteria.…”

Section: Salmonella Typhimuriummentioning

confidence: 99%

“…Preliminary studies of subunit proteins by column chromatography (27) or total proteins by one-dimensional acrylamide gel electrophoresis ( 19) have shown that protein profiles from closely related bacteria exhibit more similarities than do profiles from distantly related bacteria. In fact, 60 strains of Escherichia coli have been classified into seven groups according t o their subunit protein composition (26).…”

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confidence: 99%

Comparative Immunology of Ribosomes and Disc Gel Electrophoresis of Ribosomal Proteins from Erwiniae, Pectobacteria, and Other Members of the Family Enterobacteriaceae

Schaad

1974

International Journal of Systematic Bacteriology

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Ribosomes from erwiniae, pectobacteria, and other Enterobacteriaceae were compared by two criteria: (i) disc gel electrophoresis of 50s subunit proteins, and (ii) Ouchterlony gel diffusion of intact ribosomes. There were no significant differences in the electrophoretic protein profiles of strains of Escherichia coli, Salmonella ty phim urium, Kle bsiella pneumo niae, or En terobacter agglorn erans (synonym: Erwinia herbicola). The only signficant profile differences occurred with the faster-migrating proteins of enterobacteria grouped according to plant or animal source. Several non-enterobacteria also had similar over-all profiles; however, specific differences were evident. Immuno-diffusion data were effective in establishing relatedness at the subspecies level. A single antigenic determinant was responsible for the specificity and was found t o differ in its sensitivity t o treatment with trypsin, ribonuclease, or heat.Results of investigations on the structure and function of ribosomes suggest the possibility of using ribosomes t o determine phylogenetic relationships of bacteria. All bacterial ribosomes studied have subunits with similar sedimentation coefficients, ribonucleic acid species, and amounts of protein (28,43). Ribosomal proteins are heterogenous (13, 2 1, 46), direct products of structural cistrons (36, 37, 44) and exhibit sequence and charge similarities between closely related species (2 1, 45). On the other hand, distantly related species have ribosomal proteins of similar molecular weights (41). Thus far, however, no single method of analysis has proven completely satisfactory in expressing comparative similarities or differences.Preliminary studies of subunit proteins by column chromatography (27) or total proteins by one-dimensional acrylamide gel electrophoresis ( 19) have shown that protein profiles from closely related bacteria exhibit more similarities than do profiles from distantly related bacteria. In fact, 60 strains of Escherichia coli have been classified into seven groups according t o their subunit protein composition (26). These methods have several disadvantages: accurate, unbiased comparisons are difficult; slight variability can occur between analyses; single bands often consist of more than one protein; and it has been difficult t o infer any relationship between proteins from two different ribosomes except when they are identical. The single-band problem has been solved by the two-dimensional gel electrophoresis technique (20); however, the other problems have not. The problems associated with analysis of total or subunit proteins suggested investigating not only the individual components but also ribosomal topography. Several studies have been initiated recently t o determine the topography and intimate structural arrangement of E. coli subunit ribosomes ( 5 , 6, 9, 14, 22). Based on trypsin sensitivity and gel electrophoresis, these studies have indicated that proteins are more exposed than ribonucleic acid (RNA) in the 3 0 s subunit, with the reverse being true for th...

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Ribosomal proteins of bacterial cells: Strain- and species-specificity

Cited by 92 publications

References 14 publications

Location of Ribosomal Protein Binding Sites on 16S Ribosomal RNA

Location of Ribosomal Protein Binding Sites on 16S Ribosomal RNA

Phototrophic Green and Purple Bacteria: A Comparative, Systematic Survey

Comparative Immunology of Ribosomes and Disc Gel Electrophoresis of Ribosomal Proteins from Erwiniae, Pectobacteria, and Other Members of the Family Enterobacteriaceae

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