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Biomedical subjects

M Shani

Publications and source records attributed to M Shani.

At least 91 records · Page 5Linked to original sources

The nucleotide sequence of the rat cytoplasmic beta-actin gene.

The nucleotide sequence of the rat beta-actin gene was determined. The gene codes for a protein identical to the bovine beta-actin. It has a large intron in the 5' untranslated region 6 nucleotides upstream from the initiator ATG, and 4 introns in the coding region at codons specifying amino acids 41/42, 121/122, 267, and 327/328. Unlike the skeletal muscle actin gene and many other actin genes, the beta-actin gene lacks the codon for Cys between the initiator ATG and the codon for the N-terminal amino acid of the mature protein. The usage of synonymous codons in the beta-actin gene is nonrandom, and is similar to that in the rat skeletal muscle and other vertebrate actin genes, but differs from the codon usage in yeast and soybean actin genes.

Actins↗

Nucleotide sequence of the rat skeletal muscle actin gene.

The actins constitute a family of highly conserved proteins found in all eukaryotic cells. Their conservation through a very wide range of taxonomic groups and the existence of tissue-specific isoforms make the actin genes very interesting for the study of the evolution of genes and their controlling elements. On the basis of amino acid sequence data, at least six different mammalian actins have been identified (skeletal muscle, cardiac muscle, two smooth muscle actins and the cytoplasmic beta- and gamma-actins). Rat spleen DNA digested by the EcoRI restriction enzyme contains at least 12 different fragments with actin-like sequences but only one which hybridized, in very stringent conditions, with the skeletal muscle cloned cDNA probe. Here we describe the sequence of the actin gene in that fragment. The nucleotide sequence codes for two amino acids, Met-Cys, preceding the known N-terminal Asp of the mature protein. There are five small introns in the coding region and a large intron in the 5'-untranslated region. Comparison of the structure of the rat skeletal muscle actin gene with available data on actin genes from other organisms shows that while the sequenced actin genes from Drosophila and yeast have introns at different locations, introns located at codons specifying amino acids 41, 121, 204 and 267 have been preserved at least from the echinoderm to the vertebrates. A similar analysis has been done by Davidson. An intron at codon 150 is common to a plant actin gene and the skeletal muscle acting gene.

Actins↗

DNAase I sensitivity of genes expressed during myogenesis.

Cultures of a rat myogenic cell line were used to examine the question of whether in proliferating precursor cells genes which are programmed to be expressed later in development, in the same cell lineage, differ in DNAase I sensitivity from genes which are never expressed in these cells. Nuclei isolated from proliferating mononucleated myoblasts, differentiated cultures containing multinucleaged fibers, and rat brain, were treated with DNAase I. The sensitivity of the genes coding for the muscle-specific alpha-actin, myosin light chain 2 and the nonmuscle beta-actin was measured by blot hybridization of nuclear DNA with the corresponding cloned cDNA and genomic DNA probes. The sensitivity of these genes was compared to that of a gene not expressed in the muscle tissue. The results showed that in the muscle precursor cells, the potentiality of tissue-specific genes to be expressed is not reflected in DNAase I sensitivity. The changes which render these genes preferentially sensitive to DNAase I take place during the transition to terminal differentiation. The results showed also that the region of DNAase I sensitivity of the alpha-actin gene in the differentiated cells ends between 40 to 700 bp 5' to the structural gene. No DNAase I hypersensitive site was detected 5' to the alpha-actin gene.

Actins↗

The genes coding for the muscle contractile proteins, myosin heavy chain, myosin light chain 2, and skeletal muscle actin are located on three different mouse chromosomes.

The chromosomal distribution of murine genes expressed during differentiation of skeletal muscle cells was determined by Southern blot analysis of DNA from mouse-Chinese hamster hybrid cell lines containing incomplete subsets of mouse chromosomes. All detectable myosin heavy chain genes are located on chromosome 11. The gene for the myosin light chain 2 is located on chromosome 7. The skeletal muscle alpha-actin gene and several other actin genes, or pseudogenes, are located on chromosome 3. Additional actin DNA sequences are distributed on other mouse chromosomes.

Actins↗

Analysis of myogenesis with recombinant DNA techniques.

Recombinant phages containing the rat skeletal muscle alpha-actin gene and the cytoplasmic beta-actin gene were isolated and the structure of these genes was determined. Both genes contain a large intron in the 5' untranslated region and smaller introns at codons 41, 267 and 327. In addition, the alpha-actin contains introns at codons 150 and 204 not present in the beta-actin gene, whereas the beta-actin gene contains an intron at codon 121. The evolutionary aspects of these findings are discussed. Active genes are organized in chromatin in a conformation which renders them preferentially sensitive to digestion with nucleolytic enzymes. The DNAase I sensitivity of genes programmed to be expressed during myogenesis was tested in a cloned cell population of a myogenic cell line. It was found that these genes are not preferentially sensitive to DNAase I in the chromatin of proliferating mononucleated cells. They become DNAase I sensitive during terminal differentiation.

Actins↗

Isolation and characterization of rat skeletal muscle and cytoplasmic actin genes.

Southern blots of rat genomic DNA indicate the existence of at least 12 EcoRI DNA fragments containing actin gene sequences. By using specific probes and stringent conditions of hybridization, it was found that only one of these fragments contains sequences of the skeletal muscle alpha-actin gene. Recombinant bacteriophages originating from eight different actin genes were isolated from rat genomic DNA libraries. One of them, Act 15, contains the skeletal muscle actin gene. Another clone, Act I, contains a gene coding for a cytoplasmic actin, identified tentatively as the beta-actin gene. Both genes have a large intron very close to the 5' end of their transcribed region, followed by several small introns. DNA sequence analysis and comparison with the available data on actin genes in other organisms indicated an interesting relationship between the positions of introns and the evolutionary relatedness. Several intron sites are conserved from at least the echinoderms to the vertebrates; others appear to be present in some actin genes and not in others.

Actins↗

Skeletal muscle actin mRNA. Characterization of the 3' untranslated region.

Plasmids p749, p106, and p150 contain cDNA inserts complementary to rat skeletal muscle actin mRNA. Nucleotide sequence analysis indicates the following sequence relationships: p749 specifies codons 171 to 360; p150 specifies codons 357 to 374 together with 120 nucleotides of the 3'-non-translated region; p106 specifies the last actin amino acid codon, the termination codon and the entire 3' non-translated region. Plasmid p749 hybridized with RNA extracted from rat skeletal muscle, cardiac muscle, smooth (stomach) muscle, and from brain. It also hybridizes well with RNA extracted from skeletal muscle and brain of dog and chick. Plasmid p106 hybridized specifically with rat striated muscles (skeletal and cardiac muscle) mRNA but not with mRNA from rat stomach and from rat brain. It also hybridized to RNA extracted from skeletal muscle of rabbit and dog but not from chick. Thermal stability of the hybrids and sensitivity to S1 digestion also indicated substantial divergence between the 3' untranslated end of rat and dog skeletal muscle actins. The investigation shows that the coding regions of actin genes are highly conserved, whereas the 3' non-coding regions diverged considerably during evolution. Probes constructed from the 3' non-coding regions of actin mRNAs can be used to identify the various actin mRNA and actin genes.

Actins↗

Identification of recombinant phages containing sequences from different rat myosin heavy chain genes.

The construction and identification of a recombinant plasmid containing a cDNA insert which hybridizes specifically to myosin heavy chain mRNA is described. The plasmid was used as a probe to screen a rat genomic library for recombinant phages containing myosin heavy chain sequences. Six clones with approximately 15 k bp inserts each were isolated. Digestion with several restriction enzymes and hybridization of the fractionated DNA with the plasmid probe showed that the clones contained 3 different DNA inserts. Electron microscopy of a heteroduplex made by hybridization of DNA from two clones confirmed that the inserts originated in different genes. Hybridization of size-fractionated ECOR1 digested rat spleen DNA with the cloned probe suggested the existence of at least 5 myosin heavy chain genes.

Animals↗

Construction of recombinant plasmids containing rat muscle actin and myosin light chain DNA sequences.

The construction and partial characterization of recombinant bacterial plasmids carrying DNA sequences that hybridize with rat skeletal muscle actin and a myosin light chain mRNA is described. DNA of one clone hybridizes specifically with the muscle-specific alpha-actin mRNA. Three plasmid clones contain DNA inserts that hybridize with muscle as well as with nonmuscle actin mRNA. A fifth plasmid contains sequences complementary to mRNA coding for myosin light chain 2. DNA of this plasmid hybridizes specifically with RNA extracted from muscle and differentiated muscle cultures but not with RNA extracted from proliferating mononucleated myogenic cells.

Actins↗

Properties of simian virus 40 transcriptional intermediates isolated from nuclei of permissive cells.

A nucleoprotein complex that is an intermediate in viral transcription has been isolated from simian virus 40 (SV40)-infected BSC-1 cells after lysing infected nuclei with Sarkosyl. It contain DNA, DNA-dependent RNA polymerase II, and nascent RNA chains. RNA chain elongation continues for several hours in vitro and is dependent on exogenous ribonucleoside triphosphates. The complex sediments in neutral sucrose gradients with a main peak at about 24 to 26S. When the nascent RNA on the complex is treated with RNase A, a fraction of the RNA remains resistant to RNase and is hydrogen bonded to the DNA template. The pulse-labeled RNase-resistant RNA can be chased into RNase-sensitive RNA, indicating that it is located at the 3' terminus of the RNA chain. The rate of RNA displacement from the DNA template is consistent with an average rate of RNA chain elongation of 15 to 30 nucleotides per min. At least 70% of the RNA synthesized in this in vitro system is SV40 specific. Hybridization with the separated strands of SV40 DNA and with fragments of SV40 DNA generated with endonucleases HindII + III indicates that this RNA is complementary to all regions of the "late" SV40 DNA strand. Studies of SV40 RNA synthesis in this partially purified preparation at early and late times after infection should provide a way of locating promoter sites for transcription and identifying the form of SV40 DNA that serves as a template for late transcription.

Animals↗

The Dubin Johnson syndrome and pregnancy.

A recent summary of the scanty information about Dubin Johnson Syndrome (DJS) and pregnancy suggests that DJS is accompanied by excessive fetal mortality. This impression could not be corroborated by this study. A fetal mortality of 16% was observed in 75 pregnancies in 18 DJS patients. This value was not significantly different from the 10% mortality observed in 81 pregnancies in 15 matched control subjects. Jaundice was accentuated in 54% of pregnancies in the DJS patients and its time of onset was variable. In 6 instances jaundice was observed only following delivery.

Abortion, Spontaneous↗

Clinicoepidemiologic study of uterine cancer. Comparative aspects of the endometrial and cervical sites.

A review of all newly diagnosed cases of cervical and endometrial carcinoma in Israel during the 5-year period of 1961-1965 yielded mean annual incidence rates of 4.9/100,000 and 7.4/100,000, respectively. Cervical cancer was more prevalent in Moroccan-born women and among divorcees, while the risk of endometrial cancer was highest in older age, among the European born, and the single; it also appeared earlier in life. Postmenopausal bleeding constituted the most frequent first symptom in both sites. Fifty percent of the patients of both groups were diagnosed within 1 month, but the delay was somewhat longer in the endometrial group. Median survival was 5 years in patients with cervical cancer and above 12 years in those with cancer of the corpus. Five-year survival was 50 and 75%, respectively. Survival tended to be better in younger patients in both groups. It is expected that the gradual disappearance of intraethnic differences in Israel would lead to a decrease in the incidence of invasive cervical cancer, coupled with an increased incidence of the endometrial category.

Adult↗

Secondary pituitary insufficiency: report of three cases of ectopia or hemiagenesis of the thyroid gland.

Three patients with clinical and laboratory manifestations suggestive of pituitary insufficiency due to the presence of a pituitary tumor were found to have long-standing myxedema. The insidious appearance of the signs of hypothyroidism was explained by the cause of their disease--an ectopic thyroid gland in two patients and hemiagenesis of the thyroid in the third. Early recognition and treatment of such cases is important.

Adult↗