A mineralocorticoid-like receptor in the rainbow trout, Oncorhynchus mykiss : cloning and characterization of its steroid binding domain

Autor: Alexis Fostier, Laurent Colombe, Nicolas R. Bury, Farzad Pakdel, Yann Guiguen
Přispěvatelé: Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA)
Jazyk: angličtina
Rok vydání: 2000
Předmět:
Male
Hydrocortisone
medicine.drug_class
[SDV]Life Sciences [q-bio]
Molecular Sequence Data
Clinical Biochemistry
Biology
Biochemistry
03 medical and health sciences
Cytosol
0302 clinical medicine
Endocrinology
Mineralocorticoid receptor
Rapid amplification of cDNA ends
Complementary DNA
medicine
Animals
Humans
Testosterone
[INFO]Computer Science [cs]
Amino Acid Sequence
Cloning
Molecular
Binding site
SALMONIDAE POISSON
Molecular Biology
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
Pharmacology
0303 health sciences
Binding Sites
Base Sequence
Sequence Homology
Amino Acid
17-alpha-Hydroxyprogesterone
Organic Chemistry
DNA-binding domain
Molecular biology
Recombinant Proteins
3. Good health
Receptors
Mineralocorticoid
Mineralocorticoid
Oncorhynchus mykiss
COS Cells
Steroids
Rainbow trout
Sequence Analysis
030217 neurology & neurosurgery
Binding domain
Zdroj: Steroids
Steroids, Elsevier, 2000, 65, pp.319-328
ISSN: 0039-128X
Popis: Using reverse transcriptase polymerase chain reaction (PCR) (RT-PCR) with degenerate primers followed by 3' rapid amplification of cDNA ends PCR (3'Race-PCR) we have isolated a new fish steroid receptor cDNA sequence of 1806 bp from rainbow trout (Oncorhynchus mykiss) testis. This sequence has clear homology with various mineralocorticoid receptor cDNA sequences (rat, human, African toad: 68-70% amino acid identity), and encompasses the second part of DNA binding domain (C domain), the whole hinge region (D domain) and the steroid binding domain (E domain) plus 726 bp of 3'untranslated sequence. COS-1 cells transfected with a pCMV5 expression vector containing the whole E domain (pCMV5-rtMR) showed high affinity binding for cortisol (K(a) = 0.53+/-0.03 nM, K(d) = 1.9 nM) in the cytosol, which could not be detected in untransfected cells. Aldosterone displaced (3)H-cortisol binding, though was less effective by than unlabeled cortisol (P0.05). Competition experiments with other steroids gave the following hierarchy for the displacement of the (3) dexamethasone, whereas 17, 20beta-dihydroxy-4-pregnen-3-one and 17,20beta,21beta-trihydroxy-4 pregnen-3-one (two fish specific progestins) did not show any specific binding. These results strongly suggest that this cDNA sequence encodes a rainbow trout mineralocorticoid-like receptor, and represent the first description of such a receptor in teleost fish where aldosterone, the classic mineralocorticoid, is believed to be absent.
Databáze: OpenAIRE
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