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M Grunstein

Publications and source records attributed to M Grunstein.

At least 73 records · Page 4Linked to original sources
PubMed1987

Expression of a set of synthetic suppressor tRNA(Phe) genes in Saccharomyces cerevisiae.

Synthetic ochre and amber tRNA suppressor genes derived from the yeast tRNA(PheGAA) sequence have been constructed. They were efficiently transcribed in vitro and expressed in vivo via a synthetic expression cassette. tRNA(PheUUA) and tRNA(PheUUA) delta IVS (IVS = intervening sequence) are relatively inefficient ochre suppressors. They are toxic to the cell when expressed on a multicopy plasmid, and they do not suppress at all when present as single copies. The intron does not seem to have any effect on suppression. In contrast, the amber suppressor tRNA(PheCUA) delta IVS is efficient when expressed from a single-copy plasmid, while its efficiency is reduced on a multicopy vector.

Base Sequence↗
PubMed1986

Yeast histone H2A and H2B amino termini have interchangeable functions.

The N-terminal ends of histones H2B and H2A have very different sequences and rates of evolution. However, they both extend from the nucleosome core and are positively charged. Short sequences at the C termini of both proteins also differ from each other and appear to be hydrophilic. Deletions at the N and C termini of yeast histones H2B2 and H2A1 do not obviously affect the cell's viability under normal growth conditions. However, deletions at the N termini of both H2B and H2A in the same cell are lethal or result in greatly reduced viability. Even switching portions of the N termini between H2B and H2A to create two chimeric histone proteins within the same cell has no obvious effect on viability. This supports the argument that the N-terminal end of one protein complements the function of the other.

Amino Acid Sequence↗
PubMed1986

Sequence of glutamine synthetase from Salmonella typhimurium and implications for the protein structure.

To aid in the interpretation of the 3.5 A resolution electron density map of glutamine synthetase (GS) from Salmonella typhimurium, the nucleotide sequence of the gene coding for this enzyme has been determined. The predicted sequence of 468 amino acids (Mr = 51,628) has been compared to the sequence and sequence fragments reported by others for GS of Anabaena and Escherichia coli. The homology between the pairs of sequences is sufficiently strong to suggest that the overall three-dimensional structures of the three GS are similar. The predicted positions of alpha helices are in moderately good agreement with the electron-density map.

Amino Acid Sequence↗
PubMed1986

Albuterol protects against exercise-induced asthma longer than metaproterenol sulfate.

Both metaproterenol sulfate and albuterol are inhaled medications commonly used to prevent exercise-induced bronchospasm. Their efficacy and duration of action in controlling exercise-induced bronchospasm were compared with placebo in 18 asthmatic children (age range: 12 to 17 years) in a single-blind randomized crossover study. Standardized treadmill exercise challenges were repeated every two hours for up to six hours following the initial exercise test. With the initial exercise challenge, both active medications blocked exercise-induced bronchospasm with equal efficacy. On the other hand, when the duration of action of the medications was compared: albuterol blocked exercise-induced bronchospasm longer than metaproterenol sulfate in eight subjects, the reverse was true in only one patient, and the medications blocked for equal duration in nine subjects. Thus, although both active agents were equally efficacious in blocking exercise-induced bronchospasm initially, the duration of action of albuterol was significantly (P less than .05) longer on serial testing than that of metaproterenol sulfate. Both medications were significantly better than placebo in efficacy and duration of action.

Adolescent↗
PubMed1985

Organization, primary structure, and evolution of histone H2A and H2B genes of the fission yeast Schizosaccharomyces pombe.

The histone H2A and H2B genes of the fission yeast Schizosaccharomyces pombe were cloned and sequenced. Southern blot and sequence analyses showed that, unlike other eucaryotes, Saccharomyces cerevisiae included, S. pombe has unequal numbers of these genes, containing two histone H2A genes (H2A-alpha and -beta) and only one H2B gene (H2B-alpha) per haploid genome. H2A- and H2B-alpha are adjacent to each other and are divergently transcribed. H2A-beta has no other histone gene in close proximity. Preceding both H2A-alpha and -beta is a highly conserved 19-base-pair sequence (5'-CATCAC/AAACCCTAACCCTG-3'). The H2A DNA sequences encode two histone H2A subtypes differing in amino acid sequence (three residues) and size (H2A-alpha, 131 residues; H2A-beta, 130 residues). H2B-alpha codes for a 125-amino-acid protein. Sequence evolution is extensive between S. pombe and S. cerevisiae and displays unique patterns of divergence. Certain N-terminal sequences normally divergent between eucaryotes are conserved between the two yeasts. In contrast, the normally conserved hydrophobic core of H2A is as divergent between the yeasts as between S. pombe and calf.

Amino Acid Sequence↗
PubMed1984

Extensive purification and characterization of chromatin-bound histone acetyltransferase from Saccharomyces cerevisiae.

A strong correlation has been established between reversible acetylation of histones and transcriptional activation of chromatin. However, the function of histone acetylation remains unknown. We have approached this question by purifying histone acetyltransferase 15,000-fold from yeast and characterizing it enzymatically. Biochemical properties, including the pH and temperature optima and the Michaelis-Menten constants for both acetyl coenzyme A and histones, are similar to those reported for histone acetyltransferases from higher eukaryotes. Yeast histone acetyltransferase has a native molecular weight of 110,000 as determined by gel filtration and is tightly bound to chromatin. It displays high-substrate specificity for histones. It acetylates all four core histones in the order: H4 greater than H2B greater than H2A. 10-fold higher histone acetyltransferase activity is observed for free histones when compared to yeast polynucleosomes as a substrate.

Acetyltransferases↗
PubMed1984

Yeast may not contain histone H1: the only known 'histone H1-like' protein in Saccharomyces cerevisiae is a mitochondrial protein.

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 in yeast.

Amino Acids↗
PubMed1983

Yeast histone H2B containing large amino terminus deletions can function in vivo.

The basic amino terminus of each histone is external to the nucleosome core particle. In vitro studies have shown that the amino termini are not required for nucleosome assembly. To address the significance of these results in vivo we constructed mutations in yeast histone H2B in genetic backgrounds lacking wild-type H2B protein. We found that the protein can function in vivo even with large deletions at its amino terminus. These mutations produce no obvious phenotype and there appears to be no selection against the mutant proteins in chromatin assembly. A deletion removing a large portion of the carboxyl terminus was lethal. We conclude that much of the amino terminus of histone H2B has no essential function in vivo.

Amino Acid Sequence↗
PubMed1982

Histone H2A subtypes associate interchangeably in vivo with histone H2B subtypes.

The yeast Saccharomyces cerevisiae contains two primary sequence subtypes of histone H2B (H2B1 and H2B2) and of H2A (H2A1 and H2A2). Mutants in each of the H2B subtypes have been used to show previously that yeast cells lacking one or the other, but not both, of the H2B proteins are viable. Because H2A protein interacts in the nucleosome with H2B, we wished to determine whether specific H2A subtypes must interact with specific H2B subtypes. We describe experiments in which frameshift mutations were introduced into both of the H2A genes in vitro and the mutant genes integrated into the yeast genome, replacing the wild-type H2A genes by a subsequent recombination. Using these mutant (hta1- and hta2-) strains we find that neither H2A gene has a unique essential function during any phase of the yeast life cycle, although strains homozygous for hta1- grow more slowly. However, one functional H2A gene is required for viability because cells mutant in both H2A genes arrest at spore germination prior to bud separation. By combining these H2A mutations with the H2B mutations obtained previously, we show that all combinations of H2A and H2B subtypes produce viable cells. From these genetic experiments and electrophoretic analysis of the histone proteins of these mutants we conclude that the H2A subtypes can associate interchangeably with the H2B subtypes.

Genes↗
PubMed1982

The two yeast histone H2A genes encode similar protein subtypes.

The sequences of the two histones H2A genes in the yeast Saccharomyces cerevisiae have been determined. These genes encode two histone H2A subtypes which are 131 amino acids in length but differ at 2 amino acid positions: an Ala leads to Thr and a Thr leads to Ala change at positions 124 and 125. Thus, the two histone H2A subtypes have identical amino acid compositions. The coding regions of the two H2A genes are homologous at 369 of 393 bases (94%), with all but 2 of the 24 changes being silent. There is only 30% homology in the 5' flanking sequences of the two H2A genes. Like other eukaryotic histone genes, the yeast H2A genes are not interrupted by intervening sequences. When the yeast H2A histones are compared to those from other eukaryotes, there is at least 80% homology in amino acid sequence.

Amino Acid Sequence↗
PubMed1980

Histone H2B genes of yeast encode two different proteins.

The two genetically unlinked histone H2B genes isolated from Saccharomyces cerevisiae have been sequenced. The genes encode H2B proteins that are 130 amino acids in length and that differ by 4 amino acids. The changes betwen them are Ala leads to Ser, Lys leads to Ala, Thr leads to Val and Ala leads to Val at amino acid positions 2, 3, 27 and 35, respectively. A comparison of yeast H2B histones with those of higher eucaryotes demonstrates a high degree of homology clustered mainly at the carboxyl terminus. There is extensive base substitution between the two H2B genes in nucleotides that do not affect the amino acid sequence. DNA prelude sequences show 64% divergence. The coding regions differ at 49 of the 390 bases (12.6% divergence). 41 of these changes are in silent positions. By using the number of amino acid differences in the proteins we estimate that the two H2B genes are the result of an ancient duplication event.

Amino Acid Sequence↗
PubMed1978

Hatching in the sea urchin Lytechinus pictus is accompanied by a shift in histone H4 gene activity.

There is a distinct shift in histone mRNA synthesis at approximately 11--12 hr of sea urchin emhryogenesis, coincident with embryonic hatching. The synthesis of the blastula type (early) histone mRNAs gradually ceases at this stage and a new class of posthatching (late) histone mRNAs is produced. Briefly labeled early and late mRNAs were isolated and identified by means of RNA-DNA hybridization to different cloned histone genes. The late histone HI mRNA is approximately 40 nucleotides longer than the early HI mRNA. The H3, H2A, H2B, and H4 late mRNAs are 15--40 nucleotides shorter than their early counterparts. We present sequence evidence to show that the genes coding for the late H4 mRNA are a separate class from those that code for the early histone H4 message.

Animals↗