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M Rabinowitz

Publications and source records attributed to M Rabinowitz.

At least 73 records · Page 4Linked to original sources
PubMed1982

Species correlations between cardiac isomyosins. A comparison of electrophoretic and immunological properties.

Structural relationships between cardiac isomyosins were analyzed in 10 species using native-gel electrophoresis and radioimmunoassay. In the rat and rabbit, three types of ventricular isomyosin, V1, V2, and V3, were identified by electrophoresis. Monoclonal antibodies specific for the heavy chains of either type V1 or type V3 isomyosin in the rat and rabbit were used for comparison of immunological relationships between atrial and ventricular myosins in other species. Normal guinea pig ventricular myosin reacted with both anti-V2 and anti-V3 antibodies, but only a single myosin band was detected in this species by electrophoresis. When thyrotoxic cardiac hypertrophy was induced in guinea pigs, there was a decrease in myosin reactivity with the anti-V3 antibody and an increase in anti-V1 reactivity. This change in immunological reactivity indicated a change in proportions of two cardiac isomyosins in the guinea pig ventricle even though no myosin heterogeneity was detected by electrophoresis. In six other species including Xenopus, chicken, dog, pig, beef, and human, only a single band of myosin was detected by electrophoresis, and each myosin reacted only with the anti-V3 antibody. In the mouse, three types of ventricular myosin were also detected by electrophoresis. However, unlike V1 isomyosin of the rat and rabbit, mouse V1 isomyosin reacted equally with both anti-V1 and anti-V3 antibodies. In conclusion, we have identified highly conserved epitopes in cardiac myosin, which were found to specifically occur on either the high Ca2+-ATPase type V1 isomyosin or the lower ATPase type V3 ventricular isomyosin in most of the species examined.

Animals↗
PubMed1982

Isolation and characterization of two molecular variants of myosin heavy chain from rabbit ventricle. Change in their content during normal growth and after treatment with thyroid hormone.

We have prepared monoclonal antibodies specific for cardiac myosin heavy chain. These antibodies were used for the separation and characterization of the molecular variants of myosin heavy chain present in the rabbit heart. Two molecular forms of myosin heavy chain, HC alpha and HC beta, were isolated from the euthyroid rabbit heart by affinity chromatography. Their reactivity with our antibodies indicated that the primary structures of HC alpha and HC beta differ in at least four and share at least two antigenic determinants. Differences in the primary structure of HC alpha and HC beta were confirmed by analysis of the peptides produced by limited chymotryptic digestion of the two heavy chains. Thirteen peptide differences were consistently found. The HC alpha and HC beta variants are shown by immunologic analysis and in chymotryptic peptide profiles to be identical with the predominant forms of myosin heavy chain synthesized in the hearts of hyperthyroid and adult euthyroid rabbits, respectively. During development and maturation of the euthyroid rabbit heart, HC alpha comprises approximately 50% of the ventricular myosin between birth and 4 weeks of age; it diminishes to 20-30% by 8 weeks and to 10-20% by 12 weeks of age. Cardiac myosin from a 1-year-old rabbit is composed almost entirely of HC beta. Cardiac myosin from embryonic animals at 20 days gestation contained 20% HC alpha. These results show that HC alpha occurs normally in the euthyroid rabbit heart and that the relative proportions of HC alpha and HC beta depend on both the developmental stage and the thyroid state of the animal.

Aging↗
PubMed1982

Regulation of the nuclear-coded peptides of yeast cytochrome c oxidase.

We have analyzed the catabolite regulation of cytochrome oxidase by assaying changes in the synthesis of precursors of the nuclear-coded peptides (IV--VII) of cytochrome c oxidase in an in vitro reticulocyte cell-free system programmed with RNA isolated from cells grown in either glucose or raffinose. As a first step, we have characterized antibodies which bind to the precursors of subunits V and VI. Initial translation products for subunits IV and VII have also been tentatively identified by utilizing these antibodies. The messenger RNAs coding for the precursors of the nuclear-coded subunits fall in the expected size range of 8--15 S. Catabolite repression of the nuclear-coded oxidase peptides appears to be regulated by the abundance of their messenger RNAs. Translation of messenger RNA isolated from yeast cells grown on glucose indicates a coordinate and uniform increase in precursor synthesis during glucose derepression. In contrast, when RNA isolated from raffinose (derepressed) grown cells is used to direct cell-free translation, precursor abundance is high throughout growth, although the synthesis of some of the species changes in a complex pattern of ratio and abundance. These data indicate that the abundance of the messengers for the nuclear-coded precursors is regulated in a fashion dependent on the physiologic state of the cell.

Electron Transport Complex IV↗
PubMed1982

Analysis of transcriptional initiation of yeast mitochondrial DNA in a homologous in vitro transcription system.

We have developed an in vitro transcription system for yeast mitochondrial rRNA genes. Using highly purified yeast mitochondrial RNA polymerase and bacterial plasmids carrying DNA segments containing the mitochondrial rRNA sites of transcriptional initiation, we have been able to demonstrate correct initiation of transcription in vitro. By directly sequencing the transcription products, we show that transcription in vitro of both the 14S and 21S rRNAs is initiated at precisely the same site as it is in vivo. Transcription of the rRNA genes is highly sensitive to ionic strength and RNA polymerase concentration. Additional factors or modified conditions may be necessary to permit accurate transcription of mitochondrial protein genes.

Base Sequence↗
PubMed1982

Molecular cloning of mRNA sequences for cardiac alpha- and beta-form myosin heavy chains: expression in ventricles of normal, hypothyroid, and thyrotoxic rabbits.

We have isolated cDNA clones from thyrotoxic (pMHC alpha) and normal (pMHC beta) adult rabbit hearts. Restriction map analysis and DNA sequence analyses show that, although there is strong homology between overlapping regions of the two clones, they are distinctly different. The two clones exhibited 78-83% homology between the derived amino acid sequences and those determined by direct amino acid sequence analysis of rabbit fast skeletal muscle myosin heavy chains. The clones specify a segment of the myosin heavy chain corresponding to subfragment 2 and the COOH-terminal portions of subfragment 1. Nuclease S1 mapping was used to compare transcription of the two clones with expression of the alpha and beta forms of myosin heavy chains in the ventricles of thyrotoxic, hypothyroid (propylthiouracil-treated), and normal rabbits. Thyrotoxic ventricles contained only pMHC alpha transcripts whereas hypothyroid ventricles contained exclusively pMHC beta transcripts. These data correlate well with the presence of alpha- and beta-form myosin heavy chains. In the normal young adult rabbit, pMHC beta transcripts predominate, agreeing with the known beta form/alpha form ratio of 4:1. We therefore conclude that pMHC alpha and pMHC beta contain sequences of the alpha- and beta-form myosin heavy chain genes, respectively.

Amino Acid Sequence↗
PubMed1982

Identification of initiation sites for heavy-strand and light-strand transcription in human mitochondrial DNA.

The initiation sites for heavy (H) and light (L) strand transcription in HeLa cell mitochondrial DNA have been investigated by mapping experiments utilizing in vitro "capped" mitochondrial RNA molecules or nascent RNA chains. Mitochondrial poly(A)-containing RNA molecules were labeled at their 5' ends with [alpha-32P]GTP and guanylyltransferase ("capping" enzyme) and mapped on the mitochondrial genome by DNA transfer hybridization and S1 nuclease protection experiments. A mapping site for the capped 5' ends was found on the H strand very near to the 5' terminus of the 12S rRNA gene, and another site was found on the L strand very near to the 5' terminus of the 7S RNA coding sequence. In parallel experiments, the 5' ends of the nascent chains isolated from mitochondrial DNA transcription complexes were similarly mapped very near to the 5' termini of the 12S rRNA gene and of the 7S RNA coding sequence. The in vitro capped RNA molecules and the nascent chains thus presumably identify the same transcriptional initiation sites on the H strand and the L strand. The occurrence of a second possible initiation site for H-strand transcription 90-110 nucleotides upstream of that described above--i.e., 20-40 nucleotides upstream of the tRNAPhe gene--had been previously indicated by a mapping analysis of the nascent RNA chains and has been confirmed in the present work. The presence of two initiation sites for H-strand transcription can be correlated with other types of evidence that point to two different transcription events leading to the synthesis of a polycistronic molecule corresponding to the almost entire H strand and to the synthesis of the rRNA species.

Chromosome Mapping↗
PubMed1981

Phospholipid accumulation during the cell cycle in synchronous cultures of the yeast, Saccharomyces cerevisiae.

Phospholipid concentrations have been examined throughout successive cell cycles in synchronously growing cultures of the yeast, Saccharomyces cerevisiae. Total phospholipid phosphorus, as well as lecithin and phosphatidylethanolamine levels, exhibited stepwise increases during the cell cycle with step increments beginning just prior to new rounds of bud formation. Phosphatidylinositol and phosphatidylserine levels, on the other hand, showed what have been interpreted to be peak concentrations near the time of bud formation. Cardiolipin content varied considerably and was dependent upon the carbon source of the growth medium. Glucose-grown cells exhibited peak concentrations of cardiolipin near the time of bud formation, with marked decreases after this time. In contrast, galactose-grown synchronous cells exhibited stepwise increments in cardiolipin content, with step increases occurring near the time of new rounds of bud formation. Step or peak increases in cardiolipin, as well as all other phospholipids, were found to coincide with the time of stepwise increases in cytochrome c oxidase activity in these cells. No correlations were observed between the elaboration of mitochondrial membranes during the synchronous cell cycle and the observed patterns of phospholipid increase.

Cardiolipins↗
PubMed1981

Transcriptional initiation and 5' termini of yeast mitochondrial RNA.

We have used vaccinia virus guanylyltransferase to label polyphosphate-terminated yeast mitochondrial RNAs in vitro with [alpha-32P]GTP. Hybridization of RNA labeled in vitro indicates the presence of multiple transcriptional initiation sites in both grande and petite mitochondrial genomes. Agarose/urea gel electrophoresis of capped RNA suggests the existence of a precursor to the small (14 S) rRNA. In contrast, direct examination of the large (21 S) rRNA by partial ribonuclease T1 digestion reveals a complete lack of processing of the 5' end of the primary transcript of this RNA.

DNA Restriction Enzymes↗
PubMed1981

Mitochondrial transcription complex from Saccharomyces cerevisiae.

A DNA protein complex has been isolated from the mitochondria of Saccharomyces cerevisiae. The complex transcribes RNA complementary to mtDNA in a nonrandom manner. The RNA polymerase activity contained in the transcription complex is not dependent on the addition of exogenous template. The activity is rendered template-dependent by autolysis and can be further purified by heparin-Sepharose 4B chromatography. The activity is inhibited by heparin, Mn2+, and increasing ionic strength. The activity requires Mg2+ and ribonucleotides. The preferred template for the template dependent activity is poly[d(AT)]. The majority of the RNA synthesized by the transcription complex from endogenous DNA is complementary to the DNA strands directing the synthesis of the large and small ribosomal RNA. In yeast the 21 S and 14S rRNA genes are widely separated, therefore the transcription of these two regions but not of the intervening regions by the transcription complex suggests the existence of at least two transcriptional promoters on the yeast mitochondrial genome.

DNA, Fungal↗
PubMed1981

Purification of mitochondrial RNA polymerase from Saccharomyces cerevisiae.

The RNA polymerase from the mitochondria of Saccharomyces cerevisiae has been extensively purified by Sepharose 4B, heparin Sepharose 4B phosphocellulose, and DEAE-Sephadex A-50 chromatography. The activity co-sediments with a 45,000-dalton polypeptide at 6.3 S in glycerol gradients. The activity is inhibited by antibodies to the 45,000-dalton polypeptide. The activity is not inhibited by rifampicin or alpha-amanitin. It requires Mg2+ and is inhibited by elevated ionic strength and Mn2+. The most efficient template for the RNA polymerase is poly[d(AT)], with mtDNA being the preferred natural template. The RNA polymerase transcribes mtDNA from the petite strain F11 in a nonrandom manner.

DNA-Directed RNA Polymerases↗
PubMed1981

Molecular cloning of two fast myosin heavy chain cDNAs from chicken embryo skeletal muscle.

Recombinant DNA clones containing sequences for two different types of myosin heavy chain (HC) genes from chicken embryonic skeletal muscle were constructed and analyzed. Specificity of the clones for myosin HC was demonstrated by hybrid-arrested translation, by hybridization to a 7.0-kb mRNA, and by comparison of DNA sequences with known amino acid sequences of rabbit skeletal muscle myosin HC. Restriction enzyme and electron-microscopic heteroduplex analysis showed the presence of two distinct but homologous cDNA sequences. Hybrid melting curves indicated that both types of sequences represent fast myosin HC sequences.

Amino Acid Sequence↗
PubMed1981

CeramoMania.

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Dental Porcelain↗