TITLE:
Interaction between the Wilms tumour factor-1 element in the promoter of Amh and a downstream enhancer is required for a strong expression of the gene in pre-pubertal sertoli cells
AUTHORS:
David W. Dresser
KEYWORDS:
Mouse Cell Lines; In Vitro; Amh Promoter; SMAT; Pre-Pubertal Sertoli; Downstream Enhancer; Wilms Tumour Factor Element
JOURNAL NAME:
American Journal of Molecular Biology,
Vol.3 No.3,
July
17,
2013
ABSTRACT:
Amh (anti-Müllerian hormone)
is a single copy gene which is expressed strongly in Sertoli cells in the foetal
testis and participates in the onset of sexual differentiation. Its promoter
driving the expression of a reporter gene (d2EGFP) has been used to analyse the
role of certain defined putative elements and a downstream enhancer element
in gene expression. These experiments were carried out in vitro using a line of pre-pubertal mouse Sertoli cells,
transienly transfected with circular DNA constructs with variously mutated
promoter elements. A downstream enhancer element, situated immediately 3’ of
the polyadenylation (PA) signal for Amh, has been inserted in an equivalent
position in the d2EGFP construct. When the Amh promoter is unmodified, the downstream
enhancer (DE) is positively associated with a large increase in EGFP
expression. This is at least partly the consequence of an increased rate of expression
by individual cells. Experiments using variously truncated Amh promoters
indicate that an upstream region (-214 to -336) may play a minor role
in facilitating enhancement. However
mutation of the Wilms tumour factor-1 element, situated between the tata
box and the start of translation, results in an almost complete suppression of
enhancement.