AML1-like transcription factor induces serine elastase activity in ovine pulmonary artery smooth muscle cells.
In previous studies, we showed that induction of pulmonary artery (PA) smooth muscle cell (SMC) elastase activity by serum-treated elastin (STE) requires DNA transcription. We therefore used differential mRNA display to identify transcripts expressed coincident with elastase induction. Twenty-four individual transcripts were differentially expressed from a screen of approximately 2000 mRNA sequences. An mRNA with sequence homology to the human transcription factor AML1 was identified and subsequently cloned from ovine PA SMCs. Since AML1 binds to a consensus sequence in the promoter of neutrophil elastase, we pursued the possibility that AML1 is a candidate transcription factor for SMC elastase. We documented by immunohistochemistry that serum stimulation induces increased expression of AML1 in the nucleus of PA SMCs. We also showed that STE induction of elastase activity is associated with early expression of AML1 mRNA and protein and that AML1 consensus sequence DNA binding activity is increased in nuclear extracts of STE-treated cells. In addition, AML1 antisense oligonucleotides reduced serum induction of elastase activity. Our study thus provides the first functional evidence of AML1 transcriptional activity related to elastase genes and offers novel insights into the broader biological significance of AML1 in nonmyeloid cells.
Research Institute, The Hospital for Sick Children, and the Department of Pediatrics, University of Toronto, Ontario, Canada.
D A
DA
Wigle
K E
KE
Thompson
S
S
Yablonsky
S H
SH
Zaidi
C
C
Coulber
P L
PL
Jones
M
M
Rabinovitch
D000818:Animals; D001483:Base Sequence; D016384:Consensus Sequence; D050676:Core Binding Factor Alpha 2 Subunit; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins; D004790:Enzyme Induction; D006801:Humans; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000201:enzymology*; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins; D011651:Pulmonary Artery / Q000187:drug effects / Q000201:enzymology*; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics; D012756:Sheep; D013552:Swine; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*
D050676:Core Binding Factor Alpha 2 Subunit; D004268:DNA-Binding Proteins; D016376:Oligonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; C493728:RUNX1 protein, human; D014157:Transcription Factors; D004247:DNA; D012697:Serine Endopeptidases