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PMID: 15082777 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Protein kinase C-mediated phosphorylation of the leukemia-associated HOXA9 protein impairs its DNA binding ability and induces myeloid differentiation.

Molecular and cellular biology ·Vol. 24 ·No. 9 ·2004-05-00 ·Pages 3827-37

Vijapurkar U, Fischbach N, Shen W, Brandts C, Stokoe D, Lawrence HJ, Largman C

Abstract

HOXA9 expression is a common feature of acute myeloid leukemia, and high-level expression is correlated with poor prognosis. Moreover, HOXA9 overexpression immortalizes murine marrow progenitors that are arrested at a promyelocytic stage of differentiation when cultured and causes leukemia in recipient mice following transplantation of HOXA9 expressing bone marrow. The molecular mechanisms underlying the physiologic functions and transforming properties of HOXA9 are poorly understood. This study demonstrates that HOXA9 is phosphorylated by protein kinase C (PKC) and casein kinase II and that PKC mediates phosphorylation of purified HOXA9 on S204 as well as on T205, within a highly conserved consensus sequence, in the N-terminal region of the homeodomain. S204 in the endogenous HOXA9 protein was phosphorylated in PLB985 myeloid cells, as well as in HOXA9-immortalized murine marrow cells. This phosphorylation was enhanced by phorbol ester, a known inducer of PKC, and was inhibited by a specific PKC inhibitor. PKC-mediated phosphorylation of S204 decreased HOXA9 DNA binding affinity in vitro and the ability of the endogenous HOXA9 to form cooperative DNA binding complexes with PBX. PKC inhibition significantly reduced the phorbol-ester induced differentiation of the PLB985 hematopoietic cell line as well as HOXA9-immortalized murine bone marrow cells. These data suggest that phorbol ester-induced myeloid differentiation is in part due to PKC-mediated phosphorylation of HOXA9, which decreases the DNA binding of the homeoprotein.

MeSH Terms
Amino Acid Sequence Animals Bone Marrow Cells/cytology,physiology Casein Kinase II Cell Differentiation/physiology Cell Line DNA-Binding Proteins/metabolism Enzyme Activation Homeodomain Proteins/metabolism Isoenzymes/metabolism Leukemia, Myeloid Mice Molecular Sequence Data Myeloid Cells/cytology,physiology Phorbol Esters/metabolism Phosphorylation Protein Binding Protein Kinase C/antagonists & inhibitors,metabolism Protein Serine-Threonine Kinases/metabolism Serine/metabolism
Chemicals
DNA-Binding Proteins Homeodomain Proteins Isoenzymes Phorbol Esters homeobox protein HOXA9 Serine Casein Kinase II Protein Serine-Threonine Kinases Protein Kinase C
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Vijapurkar Ulka
Department of Medicine, University of California VA Medical Center, San Francisco, California 94121, USA.
Fischbach Neal
Shen Weifang
Brandts Christian
Stokoe David
Lawrence H Jeffrey
Largman Corey
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.24.9.3827-3837.2004
Published
2004-05-00
Pages
3827-37
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC387750
Subset
IM
Grants
NCI NIH HHS · R01 CA080029 · United States
NIGMS NIH HHS · R01 GM55814001A2 · United States
NCI NIH HHS · R01CA80029 · United States
External Links
PubMed source DOI: 10.1128/MCB.24.9.3827-3837.2004 PMC Full Text View journal details
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