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Cloning of pituitary adenylate cyclase activating polypeptide (PACAP) and preprosomatostatin I (prepro-SST I) from the brain of the gilthead seabream (Sparus aurata)

AuthorsGeba, Khaled M.; Khallaf, A. G.; Yúfera, Manuel ; Mancera, Juan Miguel; Martínez-Rodríguez, Gonzalo
Issue DateSep-2011
Citation8º Congreso de la Asociación Ibérica de Endocrinología Comparada (2011)
AbstractComplete coding sequences of gilthead sea bream (Sparus aurata) prepro-PACAP and prepro-SST-I were cloned and identified. Putative cDNA PCR fragment coding for gilthead sea bream prepro-PACAP was radioactively labeled and used as probe for the screening of a gilthead seabream brain cDNA library for prepro-PACAP positive clones. An intermediate fragment for SST-I was obtained by conventional PCR over hypothalamic cDNA. The product was cloned, sequenced, and used to obtain the full length sequence of SST-I through 3¿- and 5¿- RACE technique. Sea bream prepro-PACAP (sbPACAP) sequence consists of 2051 nucleotides (nt) and encodes a protein with 174 amino acids (aa) in its ORF. The protein starts with a 23 amino acid signal peptide, followed by a 106 aa-long PACAP-related peptide (PRP) that showed high sequence identity with PRP in other teleost fish but low similarity to mammalian GHRH (22-37%). PRP ends in a dibasic cutting site fundamental for proteolytic processing to the preproprotein to release the 45 aa mature PACAP. . Sea bream prepro-SST-I (sbPSS-I) cDNA consists of 561 nt including an open reading frame (ORF) of 365 nt, a 5¿-UTR of 26 nt and a 3¿-UTR of 169 nt containing a polyadenylation signal motif AATAAA. The ORF of the sbPSS-I cDNA encodes a 121 aa protein encompassing a putative signal peptide of 28 aa. sbPSS-I amino acid sequence comprises possible recognition sites for prohormone convertases. Changes in expression levels in juveniles acclimated to different environmental salinities (5, 12, 40 y 55¿) during 14 days were assessed. sbPACAP expression were not affected by salinity acclimation, while sbSST-I expression enhanced in specimens maintained under the highest salinity.
DescriptionTrabajo presentado en el 8º Congreso de la Asociación Ibérica de Endocrinología Comparada (AIEC), celebrado en Madrid del 5 al 7 de septiembre de 2011.
Appears in Collections:(ICMAN) Comunicaciones congresos
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