Nucleic Acids Research

Hidaka,, Yuji;Kanda,, Tadahito;Iwasaki,, Kentaro;Nomoto,, Akio;Shioda,, Tatsuo;Shibuta,, Hiroshi

doi: 10.1093/nar/12.21.7965pmid: 6095182

Abstract We determined the sequence of the 2,138 nucleotides in the Sendai virus genome just following the 3′ proximal 3,686 nucleotides which we had previously reported (Nucleic Acids Res. 11, 7317–7330, 1983). This covers the entire third gene of 1,173 nucleotides and the 3′ proximal 1,013 nucleotides of the fourth gene. Like the NP and P+C genes, both the third and fourth genes start from consensus sequence Rl (3′-UCCCAC(or UA)UUUC) at the 3′ end and the third gene terminates with consensus sequence R2 (3′-AUUCUUUUU) at the 5′ end. The third gene was identified as M, and the deduced 348 amino acids indicated that the M protein is rich in basic residues and has hydrophobic domains near the C-terminal. The fourth gene, although sequencing is not complete yet, was identified as F, since a large open reading frame found in the gene contains the characteristic sequence of 20 amino acids located at the N-terminal of the F1 protein. Analyses of the amino acid sequence suggested that the structure of the F gene product is NH2-signal peptide-F2-F1-COOH. This content is only available as a PDF. © IRL Press Limited

Certa,, Ulrich;Colavito-Shepanski,, Mary;Grunstein,, Michael

doi: 10.1093/nar/12.21.7975pmid: 6390339

Abstract It is likely that histone H1 is involved in the condensation of chromatin in eukaryotes. However, both the presence of histone H1 in yeast and the extent of yeast chromatin condensation are controversial. A 20 kD protein copurifies with yeast chromatin and was shown by other investigators to have characteristics of histone H1 protein. In an attempt to obtain a positive Identification of the 20 kD protein, we purified the protein to homogeneity and raised antibodies against it. We show here by immunofluorescence that the 20 kD protein does not localize to the nucleus but to cytoplasmic particles resembling mitochondria. Furthermore, we show by Western-blot analysis that anti-20 kD protein antibodies react to protein isolated from purified mitochondria. Finally, we present evidence based on size, charge, amino acid composition and immunological cross reactivity to suggest that the yeast 20 kD protein is likely to be the mitochondrial DNA-binding HM protein. This leaves no candidate for histone H1 1n yeast. This content is only available as a PDF. © IRL Press Limited

Mignery,, G.A.;Pikaard,, C.S.;Hannapel,, D.J.;Park,, W.D.

doi: 10.1093/nar/12.21.7987pmid: 6150463

Abstract A cDNA library from membrane-bound poly(A)+ mRNA of developing potato tubers was constructed and two classes of essentially full-length cDNA clones for a precursor to the major tuber glycoprotein, patatin were isolated. Sequence analysis shows that the two classes are approximately 98% homologous and correspond to the two major species of patatin identified previously by NH2-terminal amino acid sequence analysis. Sequence analysis also predicts that patatin 1s synthesized with a 23 amino acid signal sequence. Northern blot analysis shows that patatin mRNAs are 155O±5O nucleotides 1n length and are normally not present in polyribosomal or total RNA from stems or leaves. This content is only available as a PDF. © IRL Press Limited

McMahon, Michael, E.;Stamenkovich,, Dorothy;Petes, Thomas, D.

doi: 10.1093/nar/12.21.8001pmid: 6095183

Abstract Most of the ribosomal RNA genes of the yeast Saccharomyces cerevisiae are about 9 kilobases (kb) in size and encode both the 35S rRNA (processed to produce the 25S, 185, and 5.8S species) and 5S rRNA. These genes are arranged in a single tandem array of 100 repeats. Below, we present evidence that at the centromere-distal end of this array is a tandem arrangement of a different type of rRNA gene. Each of these repeats is 3.6 kb in length and encodes a single 5S rRNA. The coding sequence of this gene is different from that of the “normal” 55 gene in three positions located at the 3′ end of the gene. This content is only available as a PDF. Author notes *Present address: Abbott Laboratories, Abbott Park, Il 60064,USA © IRL Press Limited

Shlomai,, Joseph;Zadok,, Anat

doi: 10.1093/nar/12.21.8017pmid: 6095184

Abstract The major constituent of the trypanosomal kinetoplast ONA network are several thousand duplex DNA minicircles whose biological function is still unknown. The coding capacity and expression of these DNA minicricles, was studied in the trypanosomatid Crithidia fasciculata. Kinetoplast DNA minicircle fragments inserted into bacterial plasmid vectors were expressed in the bacterial cell. Sera elicited in rabbits, by immunization with the translational products of kinetoplast DNA minicircles in E.coli, reacted specifically with Crithidia fasciculata cellular antigens. It is inferred that kinetoplast DNA minicircles contain long open reading frames of nucleotides which are expressed in the trypanosomatid cell. This content is only available as a PDF. © IRL Press Limited

Soma,, Gen-Ichiro;Kitahara,, Namiko;Nishizawa,, Takashi;Nanami,, Hisako;Kotake,, Chikako;Okazaki,, Hideo;Andoh,, Toshiwo

doi: 10.1093/nar/12.21.8029pmid: 6095185

Abstract We have isolated a cDNA clone encoding salmon proopiomeianocortin precursor. Polyadenylated RNA was isolated from pituitary neurointermediate lobes and used to construct a cDNA library. The library was screened with 17 mer of oligodeoxyribonucleotides specific for the hexapeptide sequence in salmon β-endorphin I, Phe-Met-Lys-Pro-Tyr-Thr at positions 4–9 excluding, the third nucleotide. One positive clone, pSSM17 containing an insert of 1303 base pairs (bp) was characterized. Sequence determination revealed that it possessed sequences covering the entire regions encoding ACTH and β-lipotropin and that the mRNA had the same overall organization as those of other mammalian species, i.e., the following peptide hormones were arranged in order from 5′ upstream, ACTH including α-melanotropin and corticotropin-like intermediate lobe peptide, β-lipotropin including γ-lipotropin, β-melanotropin and β-endorphin. Amino acid sequences for putative salmon ACTH, β-, and -γ-lipotropin were predicted. Comparison of the salmon mRNA sequence with those of mammals showed that the regions of α- and β-MSH are relatively homologous, but other regions are much less so, especially in the 3′ nontranslated region where it is much longer and completely heterologous. This content is only available as a PDF. Author notes +Present address: Laboratory of Biochemistry, Seikagaku Kogyo Tokyo Research Institute, 3-1253 Tateno-cho, Higashiyamato, Tokyo 189, Japan © IRL Press Limited

Evans,, Todd;Schon,, Eric;Gora-Maslak,, Grazyna;Patterson,, Jennifer;Efstratiadis,, Argiris

doi: 10.1093/nar/12.21.8043pmid: 6095186

Abstract We have examined by fine mapping the S1 nuclease-hypersensitivity of the 5′ flanking regions of the human β-globin and rat preproinsulin II genes and of the SV40 origin/enhancer region. In all cases S1-hypersensitive sites are located in known or presumed promoter/regulatory regions. Though a consensus DNA sequence is not evident, all of these sites reside in predominantly homopurine-homopyrimidine stretches. The alternate (non-B) DNA structure which is revealed by the enzymatic probe is a sequence-dependent feature of a short stretch of DNA, which is retained upon transplantation into a foreign environment. The alternate structure exhibits S1-nicking patterns uniquely different from those associated with the presence of Z-DNA. This content is only available as a PDF. © IRL Press Limited

Cantor, Charles, R.;Efstratiadis,, Argiris

doi: 10.1093/nar/12.21.8059pmid: 6095187

Abstract S1 nuclease hypersensitive sites in the 5′ flanking regions of eukaryotic genes, present in small artificial supercoiled DNA circles, reside in homo-purine-homopyrimidine stretches. The hierarchical behavior which these sites exhibit is consistent with the notion that they act as sinks of torsional free energy. By employing DMS as a single-strand-specific reagent, we show that these sites (despite their S1 sensitivity) are regions of duplex DNA. A simple thermodynamic treatment indicates that the high torsional stress in the small DNA circles is almost certain to be relieved by the formation of alternate DNA structures. The same treatment places some constraints on the types and sizes of the regions with alternate conformation. While no definitive structural conclusions can be drawn, left-handed helices seem most consistent with the extent and the pattern of sensitivity to S1 nuclease. This content is only available as a PDF. © IRL Press Limited

Strobl, Jeannine, S.;Thompson,, E.Brad

doi: 10.1093/nar/12.21.8073pmid: 6209609

Abstract ThaI (CGCG) sites which overlap HhaI (GCGC) sites in ΦX174 and pBR322 DNA were methylated invitro with HhaI methylase and S-adenosylmethionine to yield CGmCG, or mCGCG or mCGmCG (5-methylcytosine, mC) . Methylation of either cytosine in the ThaI recognition sequence rendered the DNA resistant to ThaI cleavage. Rat pituitary cell genomic DNA was digested with ThaI or 2 other known methylation-sensitive enzymes, AvaI or XhoI. After electrophoresis and ethidium bromide staining of the DNA, all 3 enzymes showed the infrequent DNA cleavage characteristic of methylation-sensitive enzymes. Comparison of pituitary growth hormone (GH) genes bearing strain-specific degrees of methylation showed the less methylated gene to be more frequently cut by either AvaI or ThaI. ThaI resistant sites in GH genes were cleaved by ThaI after exposing cells to 5-azacytidine, an inhibitor of DNA methylation. We conclude that ThaI is a useful restriction enzyme for the analysis of mC at CGCG sequences in eukaryotlc DNA. This content is only available as a PDF. © IRL Press Limited

Valerie,, Kristoffer;Henderson, Earl, E.;deRiel, Jon, K.

doi: 10.1093/nar/12.21.8085pmid: 6095188

Abstract The denV gene from bacteriophage T4, which codes for endonudease V, a small DNA repair enzyme, has been cloned and identified by an approach combining DNA sequencing and genetics, independent of the phenotypic effect of the cloned gene. Appropriate DenV+ and DenV− deletion mutants were mapped physically to define precisely a region encompassing the denV gene. This region was sequenced in order to identify a protein-coding sequence of the correct size for the denV gene (400–500 bp). Finally, Identification was confirmed by sequencing the corresponding fragments cloned from four genetically and phenotypically well-characterized denV mutants. The denV gene 1s located at 64 kb on the T4 genome, adjacent to the ipII gene, and codes for a basic protein of 138 amino acids with a deduced molecular weight of 16,078. This content is only available as a PDF. © IRL Press Limited