Weakening of the repressive YY-1 site on the thrombospondin-1 promoter via c-Jun/YY-1 interaction

다국어 초록 (Multilingual Abstract)

Thrombospondin-1 (TSP-1) level is tightly regulated
at the transcriptional level. To determine
the detailed molecular mechanisms of TSP-1 expression,
nine serial 5'-deletion constructs of
the hum an genomic tsp-1 promoter (nucleotides
-2,220 to +756) were prepared, inserted into
luc iferase reporter plasm ids , and trans iently
transfected into the Hep3B human hepatocarcinoma
cell. Among the nine 5'-deletion constructs,
pTSP-Luc-4 (-767∼+756) had consistently
decreased luciferase activity with or without
PMA stimulation, whereas a further truncated
construct [pTSP-Luc-4' (-407∼+756)] had
increased levels of expression. By searching the
nucleotides from -767 to -407, a consensus
binding sequence (5'-CCATTTT-3') for the repressor
Yin Yang-1 (YY-1) at nucleotide -440 was
identified. The suppression induced by this site
was weakened in the presence of the region
upstream of nucleotide -767 (pTSP-Luc-1 and -2).
Nuclear protein directly bound to an oligonucleotide
containing the repressive YY-1 sequence
but the binding capacity of the sequence was
decreased by the increased c-Jun levels. Moreover,
proteins immunoprecipitated with anti-YY-1
revealed an interaction between c-Jun and YY-1
factor. These data suggest that the repressive
YY-1 site of the tsp-1 promoter could not be
functional via activating positive cis-elements on
the upstream from this site and weakened via
c-Jun/YY-1 interactions.

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