Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis最新文献

{"title":"Protection of rat liver 80 S ribosomes against ricin A chain inactivation by proteins extracted from rat liver and wheat germ ribosomal subunits with ammonium chloridemagnesium chloride","authors":"Ming-Shi Chang,&nbsp;L.L Houston","doi":"10.1016/0005-2787(81)90013-7","DOIUrl":"10.1016/0005-2787(81)90013-7","url":null,"abstract":"<div><p>Proteins extracted from wheat germ 60 S ribosomal subunits and rat liver 60 S and 40 S ribosomal subunits with <span><math><mtext>3 </mtext><mtext>M NH</mtext><msub><mi></mi><mn>4</mn></msub><mtext>Cl</mtext><mtext>75 </mtext><mtext>mM MgCl</mtext><msub><mi></mi><mn>2</mn></msub></math></span> were able to prevent the ricin A chain-mediated inactivation of untreated 80 S rat liver ribosomes. The protection of polyphenylalanine synthetic capability of 80 S ribosomes was saturable and reached 100% protection in the presence of about 20 μg of extracted protein using a uniform set of assay conditions. No protection was observed using proteins extracted from wheat germ 40 S subunits or the core fraction of rat liver 60 S subunits or protein extracted from <em>Escherichia coli</em> ribosomes or ribosomal subunits. The conclusion that the protective effect of extracted 60 S subunit proteins was specific, was further strengthened by showing that unrelated proteins such as α-lactalbumin, bovine serum albumin and lysozyme, and polypeptides such as polylysine and poly(aspartic acid), also showed no protection. If 80 S ribosomes were first treated with ricin A chain and then incubated with proteins extracted from rat liver 60 S subunits, no protection was observed. Proteins extracted with <span><math><mtext>NH</mtext><msub><mi></mi><mn>4</mn></msub><mtext>Cl</mtext><mtext>MgCl</mtext><msub><mi></mi><mn>2</mn></msub></math></span> from 60 S rat liver subunits were applied to a carboxymethylcellulose column equilibrated with 6 M urea. Stepwise elution with increasing concentrations of LiCl resulted in seven fractions. One fraction (D) contained most of the protective factor; one fraction (E) contained a lesser amount of the protective factor. Two-dimensional polyacrylamide gel electrophoresis of fraction D showed the presence of ten proteins. These data are consistent with the idea that the enzymatic target of ricin A chain is protein is nature and that fraction D contains one or more proteins that appear to act as a inhibitor against ricin A chain.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 230-242"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90013-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18298401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Differential reactivities at restriction enzyme sites","authors":"Alan D.B. Malcolm,&nbsp;John R. Moffatt","doi":"10.1016/0005-2787(81)90002-2","DOIUrl":"10.1016/0005-2787(81)90002-2","url":null,"abstract":"<div><p>A method has been developed to measure the rates of digestion by restriction enzymes at individual sites. This involves a simple arithmetical treatment of the integrated areas from a densitometer scan of an ethidium bromide stained gel. We have used this method to study the digestion by <em>Hpa</em>I, <em>Hin</em>cII and <em>Sal</em>I of pBR322 and ΦX174 DNA, and the effect of various DNA binding ligands. One of the two <em>Hpa</em>I sites in ΦX174 DNA is much more sensitive to inhibition by ligands such as netropsin, which display a preference for AT base pairs, than is the other site. Inspection of the sequences flanking the restriction sites shows that the former contains a much higher proportion of AT base-pairs than does the latter. The opposite phenomenon is observed with the two <em>Hin</em>cII sites in pBR322. This illustrates the importance of neighbouring sequences in the interaction between restriction enzymes and their cleavage sites in DNA.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 128-135"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90002-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17331955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis of a chloroplast membrane polypeptide on thylakoid-bound ribosomes during the cell cycle of Chlamydomonas reinhardii 137+","authors":"David Herrin,&nbsp;Allan Michaels,&nbsp;Eileen Hickey","doi":"10.1016/0005-2787(81)90003-4","DOIUrl":"10.1016/0005-2787(81)90003-4","url":null,"abstract":"<div><p>An in vitro protein synthesizing system consisting of thylakoid-bound polysomes supplemented with an S-100 from <em>Escherichia coli</em> has been used to investigate chloroplast membrane protein synthesis in synchronous cultures of <em>Chlamydomonas reinhardii</em>. The cell-free system produces at least one thylakoid membrane protein, D-2, on the basis of electrophoretic mobility and peptide mapping. Rough thylakoid membranes isolated in the middle and late light portion of the cell cycle synthesize substantial amounts of D-2 in vitro while the synthesis of D-2 was not detected by rough thylakoids isolated early in the light period or midway through the dark portion of the cell cycle. The same result was found when the synthesis of D-2 was investigated in vivo by pulse-labeling synchronous cell cultures with [³H]arginine. This provides further evidence for the faithful synthesis of polypeptide D-2 in vitro. These results show that one function of thylakoid-bound ribosomes is to synthesize thylakoid membrane proteins. The results also show the cell cycle regulation of membrane protein synthesis that exists within the chloroplast on thylakoid-bound ribosomes.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 136-145"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90003-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73007864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantitative aspects of the formation and loss of DNA interstrand crosslinks in Chinese hamster cells following treatment with cis-diamminedichloroplatinum(II) (cisplatin) II. Comparison of results from alkaline elution, DNA renaturation and DNA sedimentation studies","authors":"Martin F. Pera Jr.,&nbsp;Christopher J. Rawlings,&nbsp;Jason Shackleton,&nbsp;John J. Roberts","doi":"10.1016/0005-2787(81)90005-8","DOIUrl":"10.1016/0005-2787(81)90005-8","url":null,"abstract":"<div><p>The formation of interstrand crosslinks in the DNA of <em>cis</em>-diamminedichloroplatinum(II) (cisplatin)-treated Chinese hamster cells has been demonstrated by three independent, highly sensitive techniques: namely those of alkaline elution, renaturation of crosslinked DNA and alkaline sucrose gradient velocity sedimentation. Simultaneous measurement of DNA break frequency by the first two methods and a computer model combined with the third enabled the calculation of crosslink frequency after application of low doses of cisplatin. Good agreement was found between the values obtained by the three methods used here, and also with those obtained by direct measurement of the amount of crosslinked hybrid DNA in density-labelled cells treated with higher doses of cisplatin (Roberts, J.J. and Friedlos, F. (1981) Biochim. Biophys. Acta 655, 146–151). When measured by all three methods described here, crosslinking was found to increase during several hours after treatment and then to decrease with a half-life of between 12 and 24 h. For low initial levels of crosslinking, this was largely attributed to an excision repair process, since the formation of breaks in DNA was only minimal.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 152-166"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90005-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17234890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preparation and specificity of antibodies directed toward the ribose methylated nucleotide, 2′-O-methylguanosine 5′-monophosphate","authors":"Barbara S. Vold","doi":"10.1016/0005-2787(81)90017-4","DOIUrl":"10.1016/0005-2787(81)90017-4","url":null,"abstract":"<div><p>Antibodies have been produced that recognize the ribose methylated nucleotide, <span><math><mtext>2′-O-</mtext><mtext>methylguanosine</mtext></math></span> 5′-monophosphate, pGm. Specificity was tested in a radioimmunoassay. Other ribose methylated nucleotides, deoxyguanosine, and guanosine 5′-monophosphate exhibited negligible cross-reactivity. Elements of antibody recognition, in descending order of importance, were the parent base, the methylated ribose moiety, and the phosphate group.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 265-267"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90017-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17332660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acute effects of ethanol intake on albumin and total protein synthesis in free and membrane-bound polyribosomes of rat liver","authors":"J.M.G. Princen,&nbsp;G.P.B.M. Mol-Backx,&nbsp;S.H. Yap","doi":"10.1016/0005-2787(81)90001-0","DOIUrl":"10.1016/0005-2787(81)90001-0","url":null,"abstract":"<div><p>Controversial results have been reported in the last few years concerning the effects of ethanol on hepatic protein synthesis. In most of the studies no distinction has been made between the synthetic capabilities of the poly-ribosomes and the secretory product of labelled protein by the hepatocytes. In order to assess the influence of a single feeding of ethanol on the synthesis of albumin and total protein by the polyribosomes of rat liver, free and membrane-bound polyribosomes were isolated quantitatively from rats given 4–8 g ethanol per kg body weight 3–5 h before killing. The following results were obtained: (1) No difference was found in yield and size of free and membrane-bound polyribosomes isolated from control and ethanol-treated rats. The abilities to synthesize albumin and total protein were also equal for polyribosomes from both groups. (2) Addition of 1% ethanol to the incubation mixture of protein synthesis lowered albumin and total protein synthesis by 20%. No effect was observed with 0.5% ethanol. (3) Cell sap prepared from ethanol-treated rats contains a factor or factors which stimulate protein synthesis (10–15%). (4) The albumin mRNA sequence content was not changed in free and membrane-bound polyribosomal RNA fractions of ethanol-treated rats as compared to the control animals.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 119-127"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90001-0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18298400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"1H-NMR comparative studies of dinucleoside monophosphates. Influence of different factors on the conformational equilibria of nucleoside and dinucleoside monophosphates in aqueous solution","authors":"S Tran-Dinh,&nbsp;J.M Neumann,&nbsp;J Borrel","doi":"10.1016/0005-2787(81)90006-X","DOIUrl":"10.1016/0005-2787(81)90006-X","url":null,"abstract":"<div><p>In order to obtain information about different factors which exert a great influence on the conformational equilibria of mono-, di- and oligonucleotides, the average conformation of four heterodinucleoside monophosphates, ApC, CpA, ApU and UpA, was studied by ¹H-NMR at 250 MHz and compared with that of GpC, CpG, GpU, UpG and the corresponding nucleosides and nucleotides. The ratio of <span><math><mtext>N</mtext><mtext>S</mtext></math></span> conformers and the rotamer distribution of the exocyclic group were evaluated from proton-proton coupling constants, and the orientation of the base about the glycosidic bond from proton relaxation involving selective deuteration. The influence of intra- and intermolecular interactions on proton chemical shifts and conformational equilibria has been carefully studied from temperature and concentration effects. The results show that intermolecular base-base stacking does not modify the average conformation of dinucleoside monophosphates, while the intramolecular interaction (between residues of the same molecule) favours the N conformer, the <em>gg</em> rotamer and the <em>anti</em> conformation. The large upfield shift of H_5′ (lower field resonances) compared with H_5″ (higher field resonances) and the large variation of the N proportion with increasing temperature suggest that H_3′ is statistically closer to H_5′ than H_5″. There is a correlation between temperature, the <em>gg</em> rotamer and the variation in the relative chemical shifts between H_5′ and H_5″. These conclusions are in agreement with those of our previous work on GMPs using pH and temperature effects. <em>T</em>₁ relaxation studies indicate a large predominance of the <em>anti</em> conformation for both residues of ApC and ApU; these two compounds also exhibit a very high degree of association. For the naturally occurring ribonucleoside derivatives in aqueous solution, the base chemical structure, the position of the phosphate group, the temperature and pH are among the most important factors which can modify the conformation equilibria of molecules: the N conformation is generally more favoured in pyrimidine than in purine derivatives. The <em>anti</em> proportion is higher in 5′- than in 2′- or 3′-mononucleotides. However, the influence due to the position of the phosphate group is not determined in dinucleoside monophosphates. The temperature effect is negligible on the conformation of monomers but is remarkably large for the dimers: it consists of reducing the N and <em>gg</em> proportions which are preponderant in the dimers at room temperature. The pH effect is particularly large for the G residue, the conformation of which changes upon pr tonation at N⁷ (p<em>K</em> ≈ 2.3); the N conformer is favoured at acidic pH.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 167-180"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90006-X","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75577281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A computer method for construction of secondary structure from polynucleotide sequence. Possible structure of the bacterial replication origin","authors":"Tatsuo Ooi,&nbsp;Mituru Takanami","doi":"10.1016/0005-2787(81)90012-5","DOIUrl":"10.1016/0005-2787(81)90012-5","url":null,"abstract":"<div><p>A computer method to search the possible secondary structure of a long polynucleotide was developed. As a criterion for the stabilization of a secondary structure, free energy originating from base-pairings was employed, since the structure in solution would be at the free energy minimum. The method is summarized as follows: all possible helices are collected from a given nucleotide sequence under restrictions that the length of a helix is greater than <span><math><mtext>N</mtext><msub><mi></mi><mn>0</mn></msub></math></span> bases (e.g., four bases) and the free energy of the helix calculated according to free energies of two successive sequence-dependent basepairs is lower than <span><math><mtext>E</mtext><msub><mi></mi><mn>0</mn></msub></math></span> (e.g., −5 kcal/mol). The search of secondary structures of low free energy is performed by connecting one helix to another without allowing any base-pairing between loops. For connecting single-stranded regions, destabilizing free energy of 2–3 kcal/mol is added. The method was first applied to several tRNAs and the clover-leaf structure of tRNA was obtained as a free energy minimum. Then, possible secondary structures of the replication origin regions of the <em>Escherichia coli</em> and <em>Salmonella typhimurium</em> chromosomes were examined by the method, assuming that one of the strands in the origin region takes a specific secondary structure. The lowest-energy structure for the <em>E. coli</em> origin was found to be approximately identical to that for the <em>S. typhimurium</em> origin region.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 221-229"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90012-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18074094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Studies on Escherichia coli chromosome proteins II. DNA polymerases associated with the nucleoid","authors":"Tetsuhiro Moriya ,&nbsp;Kei-Ichiro Joh,&nbsp;Katsuji Hori","doi":"10.1016/0005-2787(81)90008-3","DOIUrl":"10.1016/0005-2787(81)90008-3","url":null,"abstract":"<div><p><em>Escherichia coli</em> nucleoids isolated in the presence of spermidine have been shown to contain DNA polymerase activity of which more than 95% was ascribed to DNA polymerase I and the rest to DNA polymerases II and III. It also has been found that DNA polymerases II and III or DNA polymerase III substituted entirely for DNA polymerase I or DNA polymerase I and II in the nucleoids isolated from <em>E. coli</em> mutants lacking the corresponding enzyme activities.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 189-194"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90008-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18074093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Type I DNA topoisomerases from mammalian cell nuclei interlock strands and promote renaturation of denatured closed circular PM2 DNA","authors":"Paul P. Lau ,&nbsp;Horace B. Gray Jr. ,&nbsp;Chik-Fong Wei ,&nbsp;Randy J. Legerski ,&nbsp;Donald L. Robberson","doi":"10.1016/0005-2787(81)90010-1","DOIUrl":"10.1016/0005-2787(81)90010-1","url":null,"abstract":"<div><p>Type I DNA topoisomerases from mouse ascites cell nuclei and from rat liver cell nuclei act on denatured viral closed circular PM2 DNA to produce molecules with a highly contracted structure as well as fully duplex nonsupercoiled covalently closed circular molecules. Highly contracted DNA molecules contain a novel type of topological linkage in which a strand in one region of the double-stranded molecule passes between the strands in another region of the circular molecule one or more times. Since it is also found that the action of the topoisomerase promotes renaturation of complementary strands in denatured closed circular DNA, it is suggested that formation of contracted DNA structures proceeds through renatured, duplex intermediates with highly negative superhelix densities that contain small single-stranded regions.</p></div>","PeriodicalId":100164,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis","volume":"655 2","pages":"Pages 199-209"},"PeriodicalIF":0.0,"publicationDate":"1981-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0005-2787(81)90010-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17332657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}