Molecular cloning and identification of the transcriptional regulatory domain of the goat neurokinin B gene TAC3.
J Reprod Dev
; 59(5): 463-9, 2013 Oct.
Article
in En
| MEDLINE
| ID: mdl-23812498
ABSTRACT
Neurokinin B (NKB), encoded by TAC3, is thought to be an important accelerator of pulsatile gonadotropin-releasing hormone release. This study aimed to clarify the transcriptional regulatory mechanism of goat TAC3. First, we determined the full-length mRNA sequence of goat TAC3 from the hypothalamus to be 820 b, including a 381 b coding region, with the putative transcription start site located 143-b upstream of the start codon. The deduced amino acid sequence of NKB, which is produced from preproNKB, was completely conserved among goat, cattle, and human. Next, we cloned 5'-upstream region of goat TAC3 up to 3400 b from the translation initiation site, and this region was highly homologous with cattle TAC3 (89%). We used this goat TAC3 5'-upstream region to perform luciferase assays. We created a luciferase reporter vector containing DNA constructs from -2706, -1837, -834, -335, or -197 to +166 bp (the putative transcription start site was designated as +1) of goat TAC3 and these were transiently transfected into mouse hypothalamus-derived N7 cells and human neuroblastoma-derived SK-N-AS cells. The luciferase activity gradually increased with the deletion of the 5'-upstream region, suggesting that the transcriptional suppressive region is located between -2706 and -336 bp and that the core promoter exists downstream of -197 bp. Estradiol treatment did not lead to significant suppression of luciferase activity of any constructs, suggesting the existence of other factor(s) that regulate goat TAC3 transcription.
Full text:
1
Database:
MEDLINE
Main subject:
Transcription, Genetic
/
Goats
/
Neurokinin B
/
Promoter Regions, Genetic
/
Neurons
Type of study:
Diagnostic_studies
/
Prognostic_studies
Limits:
Animals
/
Female
/
Humans
/
Male
Country/Region as subject:
Asia
Language:
En
Journal:
J Reprod Dev
Journal subject:
MEDICINA REPRODUTIVA
Year:
2013
Type:
Article
Affiliation country:
Japan