RESUMO
In this paper we describe a novel 19 kDa sarcomeric protein named telethonin. The cDNA sequence discloses an open reading frame of 167 amino acids that does not resemble any known protein. Antibodies against a recombinant telethonin fragment were used for Western blot analysis, confirming the presence of this 19 kDa protein in heart and skeletal muscle and revealing an immunofluorescence pattern typical of sarcomeric proteins, overlapping myosin. The frequency of specific cDNA clones in different libraries indicates that the telethonin transcript is amongst the most abundant in skeletal muscle. In human, telethonin maps at 17q12, adjacent to the phenylethanolamine N-methyltransferase gene.
Assuntos
Proteínas Musculares/química , Músculo Esquelético/química , Miocárdio/química , Sarcômeros/química , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , Western Blotting , Linhagem Celular , Mapeamento Cromossômico , Cromossomos Humanos Par 17/genética , Clonagem Molecular , Conectina , DNA Complementar/química , Imunofluorescência , Regulação da Expressão Gênica , Humanos , Imuno-Histoquímica , Dados de Sequência Molecular , Proteínas Musculares/genética , Músculo Esquelético/citologia , Miocárdio/citologia , RNA Mensageiro/análise , RNA Mensageiro/genética , Análise de Sequência , Transcrição Gênica/genéticaRESUMO
This work is part of the effort for sequencing chromosome XIV of Saccharomyces cerevisiae. Cosmid 14-13b contains a 37.8 kb insert derived from a partial Sau3A digestion of the genome, cloned into the BamHI site of the vector Pou6. The strategy used for sequencing is based on the fragmentation of the whole cosmid by sonication, followed by shotgun sequencing on an Applied Biosystem DNA sequencer. The clones with inserts corresponding to the vector were identified by dot-blot hybridization, without the need of sequencing. The analysis of the DNA sequence reveals 29 open reading frames (ORFs) longer than 300 bases. Nine ORFs are internal to some other ORFs. Similarity searches against DNA and protein data banks show that six ORFs correspond to already known yeast genes (OMP1, PSU1, MLS1, RPC19, DBP2, CYB5) and one ORF matches the sequence of a putative yeast gene (ESBP6).
Assuntos
Mapeamento Cromossômico , Cromossomos Fúngicos/genética , Cosmídeos/genética , DNA Fúngico/análise , Fases de Leitura Aberta , Saccharomyces cerevisiae/genética , Clonagem Molecular/métodos , Genoma Fúngico , Dados de Sequência Molecular , Análise de Sequência de DNARESUMO
In this paper the chromosomal localization of the human skeletal muscle genes Troponin-I slow-twitch (TNNI1), Troponin-I fast-twitch (TNNI2), and Troponin-C fast (TNNC2) and the refinement of the position for alpha-Tropomyosin (TPM1) and beta-Tropomyosin (TPM2) are reported. By radiation hybrid mapping, TPM1 was assigned to chromosome 15q22.1, TPM2 to chromosome 9p13.2-p13.1, TNNI1 to chromosome 1q31.3, TNNI2 to chromosome 11p15.5, and TNNC2 to chromosome 20q12-q13.11. The genomic distribution of these genes is discussed, with particular emphasis on the cluster organization of the Troponin genes.
Assuntos
Cromossomos Humanos Par 11 , Cromossomos Humanos Par 15 , Cromossomos Humanos Par 1 , Cromossomos Humanos Par 20 , Cromossomos Humanos Par 9 , Músculo Esquelético/metabolismo , Tropomiosina/genética , Troponina C/genética , Troponina I/genética , Sequência de Bases , Mapeamento Cromossômico , Primers do DNA , Humanos , Dados de Sequência Molecular , Família Multigênica , Fibras Musculares de Contração Rápida/metabolismo , Fibras Musculares de Contração Lenta/metabolismo , Reação em Cadeia da Polimerase , Tropomiosina/biossíntese , Troponina C/biossíntese , Troponina I/biossínteseRESUMO
In this paper is described the DNA sequence of cosmid 14-5 from chromosome XIV of Saccharomyces cerevisiae. The sequence is 38 855 bases long and contains 21 open reading frames (ORFs) plus three internal ORFs. Six ORFs correspond to known yeast genes (SLA2, ZWF1, BLH1, KEX2, SIN4 and URE2); two other ORFs had already been sequenced because they are adjacent to known genes; the remaining 12 ORFs are novel genes. Of these, one ORF (NII42) is particularly interesting since it shows a significant similarity to mammalian globin. Another ORF (N1254) displays two zinc finger motifs as well as a DNAJ motif.
Assuntos
Cromossomos Fúngicos/genética , DNA Fúngico/genética , Genes Fúngicos , Saccharomyces cerevisiae/genética , Animais , Cosmídeos , Proteínas Fúngicas/genética , Globinas/genética , Dados de Sequência Molecular , Fases de Leitura Aberta , Mapeamento por Restrição , Dedos de Zinco/genéticaRESUMO
A systematic study on the mRNA species expressed in the human skeletal muscle is presented in this paper. To carry on this study, a new method has been developed for the construction of unbiased cDNA libraries specially designed for the production of ESTs corresponding to the 3'-end portion of the mRNAs. The method has been applied to human skeletal muscle, where the analysis of the transcription profile is particularly difficult for the presence of several very abundant transcripts. To detect and quantify high-level mRNAs, the first 1054 ESTs were obtained from randomly selected clones. The 10 most abundant transcripts accounted for > 45% of the clones. Subsequently, these transcripts were identified by filter hybridization, thus making DNA sequencing more productive. Overall, 4370 clones were identified: 3372 by DNA sequencing and 998 by filter hybridization. The number of groups of sequences identifying individual transcripts was relatively low compared with other tissues, resulting in a total of 934 groups out of 4370 ESTs. Of these, 719 groups were represented by only one sequence.