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Biomedical subjects

H Weissbach

Publications and source records attributed to H Weissbach.

At least 109 records · Page 6Linked to original sources
PubMed1981

Chemistry and biology of E. coli ribosomal protein L12.

E. coli ribosomal protein L12, because of its unique features, has been studied in more detail than perhaps any of the other ribosomal proteins. Unlike the other ribosomal proteins that are generally present in stoichiometric amounts, there are four copies of L12 per ribosome, some of which are acetylated on the N-terminal serine. The acetylated species, referred to as L7, has not been shown, as yet, to possess any different biological activity than L12. A specific enzyme that acetylates L12 to form L7, using acetyl-CoA as the acetyl donor, has been purified from E. coli extracts. L12 is also unique in that it does not contain cysteine, tryptophan, histidine, or tyrosine, is very acidic (pI: 4.85) and has a high content of ordered secondary structure (approximately 50%). The protein is normally found in solution as a dimer and also forms a tight complex with ribosomal protein L10. There are three methionine residues in L12, located in the N-terminal region of the protein, one or more of which are essential for biological activity. Oxidation of the methionines to methionine sulfoxide prevents dimer formation and inactivates the protein. The four copies of L12 are located in the crest region(s) of the 50S ribosomal subunit. There is good evidence that the soluble factors, such as IF-2, EF-Tu, EF-G and RF, interact with L12 on the ribosome during the process of protein synthesis. This interaction is essential for the proper functioning of each of the factors and for GTP hydrolysis associated with the individual partial reactions of protein synthesis. The L12 gene is located on an operon that contains the genes for L10 and beta beta' subunits of RNA polymerase at about 88 min on the bacterial chromosome. DNA-directed in vitro systems have been used to study the unique regulation of the expression of these genes. Autogenous regulation, translational control, and transcription attenuation are regulatory mechanisms that function to control the synthesis of these proteins.

Amino Acid Sequence↗
PubMed1981

Enzymatic reduction of oxidized alpha-1-proteinase inhibitor restores biological activity.

The major serum inhibitor of proteolytic activity, alpha-1-proteinase inhibitor (alpha-1-PI), (or alpha-1-antitrypsin) can be readily inactivated by oxidation [Carp, H. & Janoff, A. (1978) Am. Rev. Resp. Dis. 118, 617-621]. This inactivation appears to be due to the oxidation of a critical methionine(s) in alpha-1-PI that is required for the inhibition of elastase activity. An enzyme from Escherichia coli that reduces methionine sulfoxide residues in protein [Brot, N., Weissbach, L., Werth, J. & Weissbach, H. (1981) Proc. Natl. Acad. Sci. USA 78, 2155-2158] can restore the biological inhibitory activity of canine oxidized alpha-1-PI.

Animals↗
PubMed1981

Enzymatic reduction of protein-bound methionine sulfoxide.

An enzyme that catalyzes the reduction of methionine sulfoxide residues in ribosomal protein L12 has been partially purified from Escherichia coli extracts. Methionine sulfoxide present in oxidize [Met]enkephalin is also reduced by the purified enzyme. The enzyme is different from a previously reported E. coli enzyme that catalyzes the reduction of methionine sulfoxide to methionine [Ejiri, S. I., Weissbach, H. & Brot, N. (1980) Anal. Biochem. 102, 393--398]. Extracts of rat tissues, Euglena gracilis, Tetrahymena pyriformis, HeLa cells, and spinach also can catalyze the reduction of methionine sulfoxide residues in protein.

Animals↗
PubMed1981

Cloning, mapping, and in vitro transcription-translation of the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase from spinach chloroplasts.

An 11.2-kilobase pair (kbp) BamHI restriction nuclease fragment from spinach chloroplast DNA has been found to contain the gene for the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase [RuP(2) carboxylase; 3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39]. The gene was located by hybridization of cloned chloroplast DNA fragments containing the maize LS gene (Bedbrook, J. R., Coen, D. M., Beaton, A. R., Bogorad, L. & Rich, A. (1979) J. Biol. Chem. 254, 905-910) to spinach chloroplast DNA cleaved with restriction nucleases. The 11.2-kbp BamHI fragment has been inserted into the BamHI site of the plasmid pBR322. The resulting recombinant plasmid, pSoe3101, was used to direct the synthesis of a protein, which was immunoprecipitable with antibody to RuP(2) carboxylase, in a partially defined in vitro transcription-translation system derived from Escherichia coli. The product synthesized in vitro has a molecular weight identical to that of authentic spinach LS. By using pSoe3101 DNA cleaved at various positions with restriction nucleases, and the in vitro transcription-translation system, the LS gene has been mapped to a 1.5-kbp region located at one end of the 11.2-kbp BamHI fragment. The direction of transcription of the LS gene on the plasmid as well as on the chloroplast chromosome has also been determined. The position of the LS gene on circular spinach chloroplast DNA is approximately 27 kbp from the start of one of the inverted repeat regions and 180 degrees from one of the rRNA-coding regions.

Journal Article↗
PubMed1980

Methylation of newly synthesized ribosomal protein L11 in a DNA-directed in vitro system.

The methylation of newly synthesized ribosomal protein L11 has been obtained in an in vitro system using lambda rifd 18 DNA as template and S-adenosyl[3H]methionine as methyl donor. About four methyl groups are incorporated per mol of L11 synthesized and the bulk of the methyl groups are present in the protein as trimethyl-lysine. lambda rifd 18 DNA also contains the genes for rRNA and newly synthesized 16 S and 23 S RNA are also methylated in this in vitro system.

Bacteriophage lambda↗
PubMed1980

Euglena gracilis chloroplast EF-Ts. Evidence that it is a nuclear-coded gene product.

Extracts of Euglena gracilis cells contain high levels of elongation factor (EF)-Ts (EF-Tschl) activity which can be assayed by measuring the rate of exchange of GDP with Escherichia coli EF-Tu . GDP. The appearance of EF-Ts activity in E. gracilis cells is light-stimulated, suggesting that the EF-Ts is required for chloroplast function. However, based on experiments with a mutant of E. gracilis lacking chloroplast DNA, as well as studies on the effect of antibiotics on EF-Ts synthesis, it is concluded that the EF-Tschl gene is nuclear-coded.

Cell Nucleus↗
PubMed1980

The effect of amino acid starvation on nucleoside uptake and RNA synthesis in Tetrahymena.

The uptake of nucleosides and the synthesis of RNA in Tetrahymena thermophila were examined following amino acid starvation. Omission of leucine, phenylalanine, or arginine from the medium resulted in a rapid decrease in the incorporation of [3H]uridine into the acid-soluble pool and acid-insoluble material (RNA). Amino acid starvation inhibited the uptake of all ribo- and deoxyribonucleosides tested but did not affect the uptake of amino acids or glucose. In addition, under the conditions used, the omission of an amino acid did not result in a large decrease in amino acid incorporation into total protein. Treatment of cells with cycloheximide or emetine gave results similar to the effects of amino acid starvation, but in these experiments the inhibition of protein synthesis was essentially complete. Nucleotide pool sizes were also measured following amino acid starvation. ATP and UTP levels were essentially unchanged, but the dTTP pool size was decreased by 40%. The decrease in RNA synthesis in vivo in the absence of an essential amino acid was reflected in the endogenous RNA synthetic activity of isolated nuclei. However, when solubilized RNA polymerase activity was measured with calf thymus DNA as template, no significant difference was observed between control and amino acid-starved cells.

Amino Acids↗
PubMed1980

RNA and protein synthesis in cultured human fibroblasts derived from donors of various ages.

RNA synthesis in human fibroblasts from donors of various ages was studied in fibroblasts made permeable to nucleoside triphosphates with the nonionic detergent Nonidet P40. Cells from donors of 11 years and older showed a 30-40% decline in total RNA synthesis. The decrease in RNA synthesis was primarily due to a lowering of RNA polymerase II activity (alpha-amanitin sensitive). Studies on the incorporation of leucine into protein also showed a 30-40% decrease in cells from older donors.

Adolescent↗
PubMed1980

L factor that is required for beta-galactosidase synthesis is the nusA gene product involved in transcription termination.

The DNA-dependent in vitro synthesis of Escherichia coli beta-galactosidase requires the presence of a soluble protein referred to as L factor [Kung, H., Spears, C. & Weissbach, H. (1975) J. Biol. Chem. 250, 1556-1562]. In the present study, comparison of physical, immunological, and biological properties shows that L factor is the product of the E. coli nusA gene. The nusA gene product is known to interact with bacteriophage lambda N gene protein and to prevent premature termination of transcription from the early lambda promoters. Our results suggest that premature transcription termination in the lac operon of E. coli may also be overcome by the nusA protein.

Bacterial Proteins↗
PubMed1980

Autogenous control of Escherichia coli ribosomal protein L10 synthesis in vitro.

The DNA-dependent in vitro synthesis of Escherichia coli ribosomal protein L10 was inhibited when L10 was added to the protein-synthesizing incubations. Addition of L10 had little or no effect on the synthesis of ribosomal protein L12, elongation factor Tu (tufB), or the beta and beta' subunits of RNA polymerase. In addition, ribosomal protein L12 did not inhibit its own synthesis or the synthesis of L10. Experiments using a mRNA-directed system showed that the inhibition of the synthesis of L10 by itself is at the level of translation of protein synthesis. The mechanism of inhibition does not appear to be due to increased degradation of L10 mRNA.

Bacterial Proteins↗