Cloning and characterization of a new cAMP responsive element binding protein on rat angiotensinogen gene
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Notice bibliographique
Résumé
Angiotensinogen (ANG) is a single peptide glycoprotein that contains 452 amino acids in human (453 amino acids in rodents). ANG releases a 10-amino acids peptide known as angiotensin l (Ang I) from it's N-terminal when acted by renin, a acidic protease. Ang I can be fiirther converted into angiotensin II (Ang II) by angiotensin converting enzyme (ACE). Ang II is one of the most potent vasoconstrictors known. Ang II also acts on the brain to mcrease blood pressure. Ang II acts on the adrenal cortex to increase the secretion of aldosterone. The levels of ANG in plasma or local tissues diïectly contribute to the levels ofAng II. Therefore the studies of regulation ofANG gene expression is important to understand the molecular mechanisms of some related diseases, such as hypertension. Previous studies m which the transfection of the fusion genes that were generated with various lengths of 5'-flanking region of the rat ANG gene linked to a bacterial chloramphenicol acetyl transferase (CAT) gene as a reporter into mouse hepatoma (Hepa 1- 6) ceUs and opossum kidney (OK) cells, identified a putative cyclic AMP responsive element (CRE) in the rat ANG gene 5'-regulatory region (N-806/-779). Compared with palindromic CRE octamer (TGACGTCA), the putative CRE of the ANG gene (ANG-CRE) is almost identical except the last two bases were in reversed order (TGACGTAC). In the present study, we isolated a fuU-length cDNA of 1345 base pairs from mouse liver cDNA library, which encodes a nuclear DNA-binding protein consisting of 436 amino acids with an apparent molecular weight of 52 kilodalton (kDa). This protein binds to the ANG-CRE, and was designated as 52-kDa protein. Southwestern blot revealed that this 52 kDa protein was present in the following tissues, such as liver, kidney, testis, brain, but not in spleen. Analysis of the deduced amino acid sequence shows no apparent basic regionleucine zipper (bZIP) stmcture, indicatmg ANG-CREB is structurally distinct fi-om bZIP family members. The antiserum against 43-kDa-CREB or ATF-2 can not interact with this 52-kDa protein, further supporting that the 52-kDa protem is immunologically different fi-om the bZIP family. The 52-kDa protein may represent a new group of CRE-binding proteins. Competitive gel mobility shift assays and Southwestern blot analysis revealed that the ANG-CREB binds to ANG-CRE, and this bindmg is not displaceable by excess amounts of CREs fi-om somatostatin (SOM), phosphoenopymvate carboxyl kmase PEPCK), and tyrosine ammo transferase (TAT) genes in vitro. These data suggest that the 52-kDa protein binds specifically to ANG-CRE and may regulate the ANG gene expression. The biological function of the 52-kDa protein is not known at present. The primary data from transient gene transfection assays showed that this protein has represser activity on the ANG gene promoter when ANG gene was stimulated by isoproterenol, but showed no effect on the ANG basal level (without any stimulation). Further experiments will be needed to study the biological function of the 52-kDa protein.
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Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
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Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découle