Ikaros stability and pericentromeric localization are regulated by protein phosphatase 1.
Ikaros encodes a zinc finger protein that is involved in gene regulation and chromatin remodeling. The majority of Ikaros localizes at pericentromeric heterochromatin (PC-HC) where it regulates expression of target genes. Ikaros function is controlled by posttranslational modification. Phosphorylation of Ikaros by CK2 kinase determines its ability to bind DNA and exert cell cycle control as well as its subcellular localization. We report that Ikaros interacts with protein phosphatase 1 (PP1) via a conserved PP1 binding motif, RVXF, in the C-terminal end of the Ikaros protein. Point mutations of the RVXF motif abolish Ikaros-PP1 interaction and result in decreased DNA binding, an inability to localize to PC-HC, and rapid degradation of the Ikaros protein. The introduction of alanine mutations at CK2-phosphorylated residues increases the half-life of the PP1-nonbinding Ikaros mutant. This suggests that dephosphorylation of these sites by PP1 stabilizes the Ikaros protein and prevents its degradation. In the nucleus, Ikaros forms complexes with ubiquitin, providing evidence that Ikaros degradation involves the ubiquitin/proteasome pathway. In vivo, Ikaros can target PP1 to the nucleus, and a fraction of PP1 colocalizes with Ikaros at PC-HC. These data suggest a novel function for the Ikaros protein; that is, the targeting of PP1 to PC-HC and other chromatin structures. We propose a model whereby the function of Ikaros is controlled by the CK2 and PP1 pathways and that a balance between these two signal transduction pathways is essential for normal cellular function and for the prevention of malignant transformation.
Popescu
Marcela
M
Department of Pediatrics, University of Wisconsin, Madison, Wisconsin 53792-4108.
Gurel
Zafer
Z
Department of Pediatrics, University of Wisconsin, Madison, Wisconsin 53792-4108.
Ronni
Tapani
T
Department of Pediatrics, University of Wisconsin, Madison, Wisconsin 53792-4108.
Song
Chunhua
C
Department of Pediatrics, University of Wisconsin, Madison, Wisconsin 53792-4108.
Hung
Ka Ying
KY
Department of Pediatrics, University of Wisconsin, Madison, Wisconsin 53792-4108.
Payne
Kimberly J
KJ
Center for Health Disparities and Molecular Medicine and Department of Pathology and Human Anatomy, Loma Linda University School of Medicine, Loma Linda, California 92350.
Dovat
Sinisa
S
Department of Pediatrics, University of Wisconsin, Madison, Wisconsin 53792-4108. Electronic address: dovat@wisc.edu.
D020816:Amino Acid Motifs / Q000502:physiology; D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D006801:Humans; D051740:Ikaros Transcription Factor / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D010766:Phosphorylation / Q000502:physiology; D017354:Point Mutation; D011485:Protein Binding / Q000502:physiology; D054645:Protein Phosphatase 1 / Q000235:genetics / Q000378:metabolism*; D055550:Protein Stability; D015398:Signal Transduction / Q000502:physiology; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism
C497239:IKZF1 protein, human; D025801:Ubiquitin; C497238:Zfpn1a1 protein, mouse; D051740:Ikaros Transcription Factor; D004247:DNA; D054645:Protein Phosphatase 1