The gene encoding the hematopoietic stem cell regulator CCN3/NOV is under direct cytokine control through the transcription factors STAT5A/B.
Cytokines control the biology of hematopoietic stem cells (HSCs) and progenitor cells in part through the transcription factors STAT5A/B. To investigate the target genes of STAT5A/B activated by cytokines in HSCs and progenitors, we performed microarray analyses using Lineage(-) Sca-1(+) c-Kit(+) (KSL) cells in the presence and absence of STAT5A/B. Stimulation with a mixture containing IL-3, IL-6, stem cell factor, thrombopoietin, and Flt3 ligand induced Ccn3/Nov mRNA over 100-fold in WT (control) but not Stat5a/b-null KSL cells. CCN3/NOV is a positive regulator of human HSC self-renewal and development of committed blood cells. Without stimulation, the Ccn3/Nov signal level was low in control KSL cells similar to Stat5a/b-null KSL cells. To determine which cytokine activates the Ccn3/Nov gene, we analyzed Lineage(-) c-Kit(+) (KL) and 32D cells using quantitative PCR and ChIP assays. Although stimulation with a mixture lacking IL-3 prevented the induction of Ccn3/Nov in control KL cells, IL-3 alone could induce Ccn3/Nov mRNA in control KL and 32D cells. ChIP assays using 32D cells revealed IL-3-induced binding of STAT5A/B to a γ-interferon-activated sequences site in the Ccn3/Nov gene promoter. This is the first report that Ccn3/Nov is directly induced by cytokines through STAT5A/B.
Kimura 木村丹香子
Akiko
A
From the Laboratory of Genetics and Physiology, NIDDK, Bethesda, Maryland 20892. Electronic address: akotalkjp@yahoo.co.jp.
Martin
Cyril
C
From the Laboratory of Genetics and Physiology, NIDDK, Bethesda, Maryland 20892.
Robinson
Gertraud W
GW
From the Laboratory of Genetics and Physiology, NIDDK, Bethesda, Maryland 20892.
Simone
James M
JM
Flow Cytometry Section, Office of Science and Technology, NIAMS, Bethesda, Maryland 20892.
Chen
Weiping
W
Microarray Core Facility with the Genomic Core Laboratory, NIDDK, Bethesda, Maryland 20892.
Wickre
Mark C
MC
From the Laboratory of Genetics and Physiology, NIDDK, Bethesda, Maryland 20892.
O'Shea
John J
JJ
Molecular Immunology and Inflammation Branch, NIAMS, National Institutes of Health, Bethesda, Maryland 20892.
Hennighausen
Lothar
L
From the Laboratory of Genetics and Physiology, NIDDK, Bethesda, Maryland 20892.
D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism* / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008817:Mice, Mutant Strains; D055514:Nephroblastoma Overexpressed Protein / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D050799:STAT5 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
C526319:CCN3 protein, human; C526320:Ccn3 protein, mouse; D016207:Cytokines; D055514:Nephroblastoma Overexpressed Protein; D012333:RNA, Messenger; D050799:STAT5 Transcription Factor; C494092:STAT5A protein, human; C494095:STAT5B protein, human; C494093:Stat5a protein, mouse; C494096:Stat5b protein, mouse; D025521:Tumor Suppressor Proteins