Biomaterial Database

Nucleotide sequence and characterization of the 5' flanking region of the rat Ha-ras protooncogene. 1987

G Damante, and S Filetti, and B Rapoport

Thyrotropin and cAMP stimulate growth of FRTL5 rat thyroid cells and increase c-Ha-ras mRNA levels. To study the mechanism by which thyrotropin enhances c-Ha-ras expression in the thyroid, we constructed a genomic library of FRTL5 DNA in the bacteriophage vector EMBL3. Using a v-Ha-ras probe (0.7-kilobase Sst I-Pst I fragment), we isolated eight clones containing portions of the FRTL5 c-Ha-ras gene. Restriction mapping of one of these clones revealed a structure very similar to that previously reported for the rat c-Ha-ras gene. We determined the nucleotide sequence of exon 1 as well as 1.15 kilobases upstream from exon 1. Blot-hybridization analysis of FRTL5 thyroid cell mRNA was performed with three DNA fragments upstream of exon 1. Two of these probes were Pst I-Pst I fragments 0.4 and 0.55 kilobases long, 1.15 and 0.6 kilobases upstream of exon 1, respectively. The third probe, a 0.6-kilobase Pst I-HindIII fragment, was immediately upstream of exon 1 and included 54 base pairs of the 5' end of exon 1. The data revealed an upstream ("-1") exon, consistent with the homology between the nucleotide sequences of our clone and the human c-Ha-ras gene in this region. Primer extension of a synthetic 30-mer oligonucleotide probe complementary to exon +1 on a FRTL5 mRNA template revealed three transcription start (cap) sites, 182, 169, and 153 bases upstream of the ATG codon. "CCAAT boxes" are present 65 and 100 bases upstream from the initial cap site. Three "GC boxes" and two C + G-rich inverted repeats characteristic of the binding site for the transcription regulation factor Sp1 are present in the 177 base pairs upstream of the initial cap site. Comparison of the rat and human c-Ha-ras -1 exons showed an area of poor homology immediately upstream of the human cap sites, followed further upstream by a region of close homology (approximately equal to 60 base pairs). The nucleotide sequence of the rat -1 exon is more similar to the v-ras sequence than is the human. The v-ras gene is truncated relative to both human and rat -1 exons.

UI	MeSH Term	Description	Entries
D009693	Nucleic Acid Hybridization	Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)	Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D011518	Proto-Oncogene Proteins	Products of proto-oncogenes. Normally they do not have oncogenic or transforming properties, but are involved in the regulation or differentiation of cell growth. They often have protein kinase activity.	Cellular Proto-Oncogene Proteins,c-onc Proteins,Proto Oncogene Proteins, Cellular,Proto-Oncogene Products, Cellular,Cellular Proto Oncogene Proteins,Cellular Proto-Oncogene Products,Proto Oncogene Products, Cellular,Proto Oncogene Proteins,Proto-Oncogene Proteins, Cellular,c onc Proteins
D011519	Proto-Oncogenes	Normal cellular genes homologous to viral oncogenes. The products of proto-oncogenes are important regulators of biological processes and appear to be involved in the events that serve to maintain the ordered procession through the cell cycle. Proto-oncogenes have names of the form c-onc.	Proto-Oncogene,Proto Oncogene,Proto Oncogenes
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D004262	DNA Restriction Enzymes	Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.	Restriction Endonucleases,DNA Restriction Enzyme,Restriction Endonuclease,Endonuclease, Restriction,Endonucleases, Restriction,Enzymes, DNA Restriction,Restriction Enzyme, DNA,Restriction Enzymes, DNA
D005091	Exons	The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA.	Mini-Exon,Exon,Mini Exon,Mini-Exons
D005796	Genes	A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.	Cistron,Gene,Genetic Materials,Cistrons,Genetic Material,Material, Genetic,Materials, Genetic
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000242	Cyclic AMP	An adenine nucleotide containing one phosphate group which is esterified to both the 3'- and 5'-positions of the sugar moiety. It is a second messenger and a key intracellular regulator, functioning as a mediator of activity for a number of hormones, including epinephrine, glucagon, and ACTH.	Adenosine Cyclic 3',5'-Monophosphate,Adenosine Cyclic 3,5 Monophosphate,Adenosine Cyclic Monophosphate,Adenosine Cyclic-3',5'-Monophosphate,Cyclic AMP, (R)-Isomer,Cyclic AMP, Disodium Salt,Cyclic AMP, Monoammonium Salt,Cyclic AMP, Monopotassium Salt,Cyclic AMP, Monosodium Salt,Cyclic AMP, Sodium Salt,3',5'-Monophosphate, Adenosine Cyclic,AMP, Cyclic,Adenosine Cyclic 3',5' Monophosphate,Cyclic 3',5'-Monophosphate, Adenosine,Cyclic Monophosphate, Adenosine,Cyclic-3',5'-Monophosphate, Adenosine,Monophosphate, Adenosine Cyclic
D000595	Amino Acid Sequence	The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.	Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein