المؤلفون: Susan E. Stred, Joseph L. Messina

المصدر: Molecular and cellular endocrinology. 204(1-2)

مصطلحات موضوعية: Male, DNA, Complementary, Biology, Biochemistry, Rats, Sprague-Dawley, chemistry.chemical_compound, Endocrinology, Hemopexin, Complementary DNA, Sequence Homology, Nucleic Acid, Gene expression, Tumor Cells, Cultured, Coding region, Animals, Northern blot, RNA, Messenger, Molecular Biology, Heme, Gene Library, Messenger RNA, cDNA library, Molecular biology, Rats, body regions, chemistry, Gene Expression Regulation, Liver, Growth Hormone

الوصف: A cDNA library from the liver of a growth hormone (GH)-treated hypophysectomized rat was constructed and screened for GH-inducible genes (GIGs). Three cDNAs specific for putative GIG mRNAs (GIG-3, -7 and -12) were isolated and, when sequenced, were found to be homologous to portions of rat hemopexin, a Class 2 acute-phase gene. Hemopexin is an essential heme scavenger produced primarily in the liver, which upon binding to free heme, transports it to the liver where the heme iron is re-utilized. Hemopexin has not been previously described as being GH-responsive. GIG-3 and GIG-12 encode overlapping portions of the entire coding sequence starting within a few hundred base pairs from the 5' end of the hemopexin mRNA, and GIG-7 encodes the 3'-most end of the hemopexin mRNA. Northern analysis and ribonuclease protection assays of RNA from livers of control rats using the cDNA probes demonstrated a major transcript of approximately 2.0 kb. The hemopexin mRNA was low or undetectable in livers of hypophysectomized rats. Daily treatment with bovine growth hormone (bGH) for 10 days restored hemopexin mRNA to levels comparable or greater than that of intact rats. GH-dependence in cultured rat H4IIE hepatoma cells was then examined. Using hemopexin cDNA probes (GIG-3, -7, and -12) we identified a mRNA on Northern blots, which increased in concentration following bGH, compared with untreated cells. When measured by ribonuclease protection assay, a maximal increase in hemopexin mRNA concentration was obtained following 4-6 h of bGH administration. We conclude that hemopexin is a GH-inducible gene in rat liver in vivo and in cultured rat hepatoma cells.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::351e0f65bf7b04bdfeb04c74b579464fTest
https://pubmed.ncbi.nlm.nih.gov/12850285Test

المؤلفون: Carla Berard, Shigeho Ijiri, John M. Trant

المصدر: Molecular and cellular endocrinology. 164(1-2)

مصطلحات موضوعية: Untranslated region, DNA, Complementary, Molecular Sequence Data, Biochemistry, Evolution, Molecular, Exon, Endocrinology, Aromatase, Complementary DNA, Stingray, Coding region, Animals, Amino Acid Sequence, Molecular Biology, Genetics, biology, Base Sequence, cDNA library, Fishes, biology.organism_classification, Molecular biology, Organ Specificity, COS Cells, biology.protein, Dasyatis sabina, Sequence Alignment

الوصف: Cytochrome P450 aromatase (P450arom; CYP19) mediates the conversion of androgens to estrogens and its activity has been found in all vertebrates studied to date. This study describes the full-length cDNA encoding the ovarian form of P450arom and the differences in the 5'-untranslated region (5'-UTR) of the extra-gonadal P450arom transcript expressed by the Atlantic stingray (Dasyatis sabina). Elasmobranchs (cartilaginous fishes such as sharks, rays and skates) diverged from the other vertebrates more than 350 million years ago, therefore the stingray P450arom cDNA may represent an ancient form of this gene. Northern blot analysis showed that the ovarian follicle expressed transcripts of 3.1 and 1.7 kb in size which correspond to the clones isolated from a stingray ovarian follicle cDNA library. Both transcripts consisted of an identical 1.5 kb coding region and a 41 bp 5'-UTR, however the 3'-UTRs differed in the use of the most proximate and the most distal of four polyadenylation signals. COS cells transfected with the 1.7 kb cDNA had twice the aromatase activity as cells transfected with the 3.1 kb cDNA. The coding region of the cDNA predicted a 58.5 kDa protein which consisted of 511 residues. Alignment of the stingray protein indicates that the P450arom is equally identical (53-59%) to all other vertebrate forms of P450arom characterized to date, thus indicating a common ancestry. The evolutionary relationship of the stingray form of P450arom clearly predates the other forms and belongs to a unique lineage. Transcripts of P450arom were expressed in ovarian follicles (of all sizes), the testis, the pituitary, in all sections of the brain, and in the kidney. The extra-gonadal transcripts appear to encode a protein identical to the ovarian form, however, the 5'-UTR was 657 bp longer presumably due to the transcription of an untranslated 'first exon' as seen in the mammalian form of this gene.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::96c7acd5899a6bf31608e80d338c6bf0Test
https://pubmed.ncbi.nlm.nih.gov/11026568Test

المؤلفون: Chihiro Azuma, Fumitaka Saji, Tadashi Kimura, Kazumasa Hashimoto, Yuji Murata, Yoshihiro Tokugawa, Katsutoshi Nobunaga, Yasue Kubota

المصدر: Molecular and cellular endocrinology. 124(1-2)

مصطلحات موضوعية: DNA, Complementary, Transcription, Genetic, TATA box, Molecular Sequence Data, Gene Dosage, Biology, Biochemistry, Exon, Mice, Endocrinology, Rapid amplification of cDNA ends, Pregnancy, Complementary DNA, Coding region, Animals, Amino Acid Sequence, RNA, Messenger, Cloning, Molecular, Promoter Regions, Genetic, Molecular Biology, Gene, Peptide sequence, Base Sequence, Sequence Homology, Amino Acid, Uterus, Intron, Exons, Sequence Analysis, DNA, Molecular biology, Introns, Gene Expression Regulation, Genes, Receptors, Oxytocin, Female

الوصف: To determine the structure of the mouse oxytocin receptor (OTR) gene, we have screened a mouse genomic library and confirmed cDNA sequence with rapid amplification of cDNA ends. Southern blot using human OTR cDNA probes indicated that the mouse genome has a single copy of the gene. The predicted amino acid sequence is 91% identical to human OTR. The gene contains 4 exons and 3 introns. Exons 1 and 2 contain the 5′ untranslated region, with exons 3 and 4 encoding the amino acids of the receptor. Intron 3 interrupts the coding region at the same location, after transmembrane domain 6, as in OTR genes of other species. The promoter region lacks an apparent TATA box but contains multiple putative interleukin-response elements and estrogen responsive elements. Expression of OTR gene in the uterus during pregnancy reached maximum at day 20 of gestation. The information obtained from the mouse OTR gene facilitates further comparative and biochemical analysis for protein structure-function relationships and gene regulatory mechanisms.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::cae87e32bfa7bb707cd5cb1d4a1a1f88Test
https://pubmed.ncbi.nlm.nih.gov/9027321Test

المؤلفون: Kirk G. Peterson, Janet F. Noble, Michael J. McPhaul, Fredrick W. George, Edwin D. Lephart

المصدر: Molecular and cellular endocrinology. 70(1)

مصطلحات موضوعية: Male, Molecular Sequence Data, Restriction Mapping, Heme, Biology, Biochemistry, Endocrinology, Aromatase, Complementary DNA, Sequence Homology, Nucleic Acid, Tumor Cells, Cultured, Coding region, Animals, Humans, Northern blot, Amino Acid Sequence, RNA, Messenger, RNA Processing, Post-Transcriptional, Molecular Biology, Messenger RNA, Base Sequence, cDNA library, Ovary, Intron, DNA, Exons, Blotting, Northern, Molecular biology, Rats, biology.protein, Neoplastic cell, Female, Leydig Cell Tumor

الوصف: The conversion of androgens to estrogens is catalyzed by a complex of enzymes that includes a specific cytochrome P-450 aromatase (P-450arom). In this paper we describe the high level expression of aromatase activity in the rat Leydig cell tumor line, R2C. We also report the isolation of cDNA clones encoding the rat aromatase P-450arom from a cDNA library prepared from this cell line. Analysis of these cDNA clones predicts a protein sequence with a high degree of sequence conservation when compared to the chicken and human P-450arom enzymes. Notably, four of the cDNA clones were found to lack the last coding exon that contains the heme-binding domain, a structural feature essential for aromatase activity. These clones were found to contain instead a segment of genomic DNA derived from an unspliced intron. Northern analysis using a fragment of the coding region of the rat P-450arom cDNA as probe revealed that three species of P-450arom mRNa are expressed in rat ovary that are similar to those identified in RNA samples prepared from the rat R2C cell line. Analysis of the same samples of RNA using a probe derived from the 3' terminal intron segment of the rat aromatase cDNA clones that lack the heme-binding domain indicates that two of the species of aromatase mRNA transcripts present in both rat ovary and R2C cell lack the heme-binding domain and thus must encode a nonfunctional aromatase protein. These findings have important implications for the measurement of aromatase mRNA and appear to explain why three sizes of rat P-450arom mRNA exist on Northern analysis and why previous studies failed to demonstrate a clear relationship between aromatase mRNA, protein, and enzymatic activity in the rat ovary.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bb7cf1db3cad33c1caa7e02c3c20083bTest
https://pubmed.ncbi.nlm.nih.gov/2340950Test

المؤلفون: Jacqueline Van Sande, Gilbert Vassart, Jacques Emile Dumont, Carine Maenhaut, Marc Parmentier, Anne Lefort, Frédérick Libert, Jason Perret, Catherine Gerard

المصدر: Molecular and cellular endocrinology. 68(1)

مصطلحات موضوعية: Genetics, Untranslated region, cDNA library, Molecular Sequence Data, Genetic Variation, Receptors, Thyrotropin, DNA, Biology, Biochemistry, Molecular biology, Thyrotropin receptor, Exon, Endocrinology, Dogs, Complementary DNA, Coding region, Animals, Amino Acid Sequence, Chromosome Deletion, Cloning, Molecular, Receptor, Molecular Biology, Gene, Repetitive Sequences, Nucleic Acid

الوصف: A clone coding for a variant form of thyrotropin receptor was isolated from a dog thyroid cDNA library. It was characterized by a 75 bp deletion in the coding region, additionally to minor modifications in the 3' untranslated region. The corresponding 25 amino acids deletion is located in the long NH2 terminal extracellular domain which is characteristic of the glycoprotein hormone receptors. This region of the protein is composed of imperfect repeats and the deletion corresponds exactly to one of the repeat units. This suggests that the repeats correspond to individual exons in the thyrotropin receptor chromosomal gene. It is not known whether the deletion of the repeat and the concomitant suppression of one of the N-glycosylation sites of the molecule do alter the receptor function.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::77fcd09ce478fbdb32fea89e1aea8724Test
https://pubmed.ncbi.nlm.nih.gov/2303156Test

المؤلفون: Hirai Toshiaki, Takikawa Hiroo, Kato Yukio

المصدر: Molecular and cellular endocrinology. 63(1-2)

مصطلحات موضوعية: Untranslated region, Swine, Molecular Sequence Data, Thyrotropin, Biology, Biochemistry, Endocrinology, Complementary DNA, Coding region, Animals, Amino Acid Sequence, Cloning, Molecular, Protein Precursors, Molecular Biology, Peptide sequence, chemistry.chemical_classification, Messenger RNA, Base Sequence, cDNA library, Nucleic acid sequence, DNA, Molecular biology, Amino acid, chemistry, Glycoprotein Hormones, alpha Subunit

الوصف: cDNA clones encoding precursors of glycoprotein hormone common alpha-subunit (pre-alpha) and of thyroid stimulating hormone beta-subunit (pre-TSH beta) were isolated from a porcine anterior pituitary cDNA library using DNA probes, and the nucleotide sequences were determined. The nucleotide sequence of pre-alpha cDNA contained an entire coding region (360 bases) including 5' and 3' untranslated regions. The pre-alpha mRNA was about 900 bases long. The predicted amino acid sequence consisted of a signal peptide of 24 amino acid residues and a mature alpha-subunit protein of 96 residues. Six amino acid residues at the amino terminus of the predicted mature protein had not been found by direct amino acid sequencing of the purified protein. The nucleotide sequence of pre-TSH beta cDNA contained an entire coding region and a 3' untranslated region which has two polyadenylation signals. The length of the pre-TSH beta mRNA was about 500 bases long. The predicted amino acid sequence consisted of a signal peptide of 20 amino acid residues, a mature protein of 112 residues and an additional extension of six amino acid residues at the carboxyl terminus, which had not been found in the amino acid sequence of the purified protein. The coding sequences of the cDNAs showed high homologies with those of other mammalian species (84-93% for pre-alpha and 81-94% for pre-TSH beta). Comprehensive data of our serial molecular cloning for porcine glycoprotein hormones revealed low but significant homologies (34-40%) among three beta-subunits. Upon comparison of frequency of (U)n A sequence in 3' untranslated region, porcine pre-alpha and pre-TSH beta mRNAs were grouped into a moderate class of mRNA stability whereas porcine pre-FSH beta and pre-LH beta were grouped into unstable and stable classes, respectively.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f7b981d3f0e6ee3542c8d3c9cb1a0db9Test
https://pubmed.ncbi.nlm.nih.gov/2473932Test

المؤلفون: Christine A. Weaver, David F. Gordon, Byron Kemper

المصدر: Molecular and cellular endocrinology. 28(3)

مصطلحات موضوعية: Sequence analysis, TATA box, DNA, Recombinant, Biology, Biochemistry, Endocrinology, Complementary DNA, Consensus sequence, Escherichia coli, Coding region, Animals, RNA, Messenger, Cloning, Molecular, Molecular Biology, Gene, Messenger RNA, Base Sequence, Nucleic acid sequence, Nucleic Acid Hybridization, RNA-Directed DNA Polymerase, DNA, DNA Restriction Enzymes, Molecular biology, Parathyroid Hormone, Cattle, Plasmids

الوصف: The sequence of bovine parathyroid hormone mRNA has been determined by sequence analysis of near full-length cloned DNA complementary to the mRNA. Restriction fragments hybridized to the mRNA and extended toward the 5' terminus with reverse transcriptase were analyzed to derive the sequence not present in cDNA. The reverse transcripts were heterogeneous in length with three major stopping points within 8 nucleotides of each other and a minor stop about 30 bases further toward the 5' terminus of the mRNA. The sequence of the gene corresponding to the minor reverse transcript begins with the sequence 5' XXXATATATAAAA which contains the consensus sequence for a TATA box, a putative eukaryotic promoter sequence. Assuming that the major reverse transcriptase stop nearest the 5' terminus of the mRNA, which is 24 bases downstream from the TATA box, represents the beginning of bovine PTH mRNA, the mRNA contains 672 nucleotides, 100 in the 5' noncoding region, 348 in the coding region and 224 in the 3' noncoding region. Bovine PTH mRNA contains 38% G and C bases. The 3' noncoding region is particularly rich in A and U bases with the last 100 nucleotides of the molecules containing 46% U and 32% A. As with other mRNAs, the sequences CO and UAG occur much less than expected. The 5' noncoding region does not contain an AUG before the initiator codon and contains two potential regions that could base-pair with sequences near the 3' terminus of 18S ribosomal RNA. The sequence AAUAAA is present 14 nucleotides from the polyadenylic acid at the 3' terminus. Bovine PTH mRNA exhibits extensive homology with human PTH mRNA.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c64154b19b3913c8387a8d4ed31601ccTest
https://pubmed.ncbi.nlm.nih.gov/6185374Test