RESUMEN
Overlapping cDNA clones for the smaller son gene transcript were isolated by several steps of human placenta cDNA library walking. Here we report the structure of repeats that exist at the long open reading, present in the structure of this smaller (5.6 kb) transcript of human gene son. We were able to identify in this transcript four areas of complete tandem repeats, formed by several steps of duplication. For three of them the extent of homology between amino acid sequences of repeated elements is higher than between their nucleic acid sequences. It means that amino acid sequences of repeated elements are conserved, that could reflect the structural significance for the function of hypothetical protein product of human gene son. On the basis of the results of analysis we proposed the model for the formation of repeats by processes of duplication, mutation and natural selection pressure at the expressed protein level.
Asunto(s)
Secuencias Repetitivas de Ácidos Nucleicos , Transcripción Genética , Secuencia de Aminoácidos , Secuencia de Bases , Codón , ADN , Humanos , Datos de Secuencia Molecular , Sistemas de Lectura AbiertaRESUMEN
We have investigated the functional activity of human son gene, that possesses the homology to mos and myc genes. Specific antibodies (antiserum) were raised to synthetic peptide, that corresponds to son-protein 943-963 amino acid residues. With this antiserum the presence of son-protein was showed in lysates of cultured human cells transformed by adenovirus type 5, RAT 2 cells and primary human embryonic fibroblasts. son-Protein molecular weight (92 kDa) was determined by the method of electrophoresis in SDS-polyacrylamide gel. Thus, it was shown the presence of son gene protein in animal and human cells. To determine a possible son gene role in mammalian cells we have cloned the 3' part (2667 b.p.) of son cDNA in retroviral vector pPS-3-neo. Transformed cells of different lines were selected. A large portion of this cells changed their morphology. New protein product (120 k), that reacted with antiserum to son specific peptide, was found together with p92son in these clones.
Asunto(s)
Transfección , Células 3T3 , Adenoviridae , Secuencia de Aminoácidos , Animales , Línea Celular Transformada , Transformación Celular Viral , Electroforesis en Gel de Poliacrilamida , Humanos , Mamíferos , Ratones , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , RatasAsunto(s)
Genes ras/efectos de los fármacos , Oligonucleótidos Antisentido/farmacología , ARN Mensajero/efectos de los fármacos , División Celular/efectos de los fármacos , Células Cultivadas , Embrión de Mamíferos , Fibroblastos/efectos de los fármacos , Genes ras/genética , Humanos , Oligonucleótidos Antisentido/genética , ARN Mensajero/genética , Factores de Tiempo , Células Tumorales Cultivadas/efectos de los fármacos , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
AO region (884 b.p.) of ORAgp5 locus has been sequenced and proved to contain mos-related regions, as well as fragments structurally similar to proretroviral elements and Alu repeats. The data obtained are in accordance with earlier hypotheses on retroviral involvement in mos gene family generation and the role of Alu repeat insertion in the inactivation of mos-related genes. Molecular hybridisation studies showed the structural conservation of the segment in ORAgp5 locus comprising the AO region among higher primates. These data indicate that AO region of ORAgp5 was formed not later than 65-70 MYR ago and that the present structure of this region was kept during last 50 MYR.
Asunto(s)
Secuencia de Bases , Genes Virales , Genes , Oncogenes , Retroviridae/genética , Homología de Secuencia de Ácido Nucleico , Clonación Molecular , Humanos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Mapeo RestrictivoRESUMEN
From the human embryonic cDNA library a son3 transcript was cloned and sequenced (1454 base pairs). Determination of its sequence revealed only one open reading frame, whereas five other frames contained a number of termination codons. Translation of the son3 sequence in the unique open reading frame into the amino acid sequence by computer and comparison with the NBRF protein data bank showed that the son3 fragment codes for a new previously unknown polypeptide with the following properties: a) it contains a cluster of short tandemly arranged repeats 7-12 amino acid in length located in the middle part of son3; b) it comprises a region homologous to DNA binding structural proteins (for example, gallin, 55%) and to regulatory proteins coded by the family of proto-oncogene myc; c) it comprises a region homologous to the oncoprotein coded by proto-oncogene mos (human, murine).
Asunto(s)
Secuencia de Bases , Proteínas de Unión al ADN/genética , Homología de Secuencia de Ácido Nucleico , Secuencia de Aminoácidos , Clonación Molecular , ADN/genética , Humanos , Datos de Secuencia Molecular , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas/genética , Secuencias Repetitivas de Ácidos NucleicosRESUMEN
Previously described genomic sequences, related to viral oncogene mos (human mos pseudogene gp5 and main c-mos gene), are either totally silent or show highly limited degree of expression in some specialized tissues. The mos-related clone K51 isolated previously from rat genome was used here to study expression of the gene comprised. We have demonstrated the presence of abundant transcripts in rat and human brain, lung, skin and muscle. The pattern of transcription changes after the birth and is shown to depend on the presence of higher molecular weight transcripts in adult tissues. With the help of hybridization to K51 probe, we have isolated three clones from human embryonic cDNA library. One of these (son3) also hybridizes to the viral mos gene. These results evidence strongly in favour of the hypothesis that K51 recombinant contains a new member (son) of mos-related family of oncogenes, with the broad spectrum of tissue specificity.
Asunto(s)
Familia de Multigenes , Transcripción Genética , Animales , Clonación Molecular , ADN/genética , Embrión de Mamíferos , Regulación de la Expresión Génica , Humanos , Hibridación de Ácido Nucleico , Especificidad de Órganos , Proto-Oncogenes , ARN/genética , ARN/aislamiento & purificación , RatasRESUMEN
The occurence of members of mos oncogene family in the vertebrates genome has been studied with the help of highly labeled single-stranded DNA probes. These included subgenic v-mos clones as well as the unique sequence--specific recombinants from mos-related human locus gp5 and the K51 locus from rat genome. The probe from gp5 (mos pseudogene) interacts only with DNA of primates and of rodents. On the other hand, mos gene and the gene from K51 locus are present in all vertberates tested. Recent duplication of the main mos gene in Artiodactyla and Perissodactyla orders of mammals was identified. The persistence of K51 and mos genes during evolution indicates their importance. The segregation of three mos-related genes in human-hamster hybrids points to their location on different human chromosomes.