MULTIPLE BINDING-SITES FOR NUCLEAR FACTORS IN THE 5'-UPSTREAM REGION OF 2 ALPHA(2U)-GLOBULIN GENES - IMPLICATIONS FOR HORMONE-REGULATED ANDTISSUE-SPECIFIC CONTROL
P. Vandijck et al., MULTIPLE BINDING-SITES FOR NUCLEAR FACTORS IN THE 5'-UPSTREAM REGION OF 2 ALPHA(2U)-GLOBULIN GENES - IMPLICATIONS FOR HORMONE-REGULATED ANDTISSUE-SPECIFIC CONTROL, Journal of steroid biochemistry and molecular biology, 45(5), 1993, pp. 353-366
In order to understand the tissue- and hormone-specific control of alp
ha2u-globulin synthesis we isolated the 5'-upstream putative regulator
y region of two alpha2u-globulin genes: RAP 01 and RAO 01. Both clones
seem to be expressed in rat liver. DNAseI footprinting analysis after
incubation with rat liver nuclear extracts was used to identify regio
ns of potential interest. Particular attention was paid to protected r
egions located in the neighbourhood of domains which, according to our
previous studies, interact specifically with androgen- and glucocorti
coid-receptor complexes. Fifteen DNAseI footprints could be mapped in
clone RAO 01 (bp - 758 up to the cap site). Nineteen footprints were o
bserved in the corresponding region of RAP 01. Differences in the foot
printing patterns were mainly observed in the more distal regions. Our
data confirm the presence in both clones of two binding sites for the
liver enriched factor pseudo-NF1 and one site for C/EBP previously ob
served in other alpha2u-globulin genes. In addition we have been able
to demonstrate, in RAP 01 only, a binding site for transferrin-liver f
actor 1. No differences in footprinting patterns could be demonstrated
using liver nuclear extracts derived from animals with a high hepatic
expression of alpha2u-globulins (normal male rats) and animals with l
ow to absent expression (prepubertal rats, female rats, rats with the
testicular feminization syndrome, diabetic rats and hypophysectomized
animals). Transfection experiments indicate that a fragment of RAP 01
(bp - 643 up to - 617) is able to act as a glucocorticoid and as an an
drogen response element. Larger fragments of RAP 01 and fragments of R
AO 01 are ineffective. It is concluded that the expression of individu
al alpha2u-globulin genes is probably the result of combinatorial inte
ractions of several trans-acting factors with appropriate cis-acting e
lements. Moreover, important sites for tissue-specific and hormone-reg
ulated expression may be situated outside the regions investigated.