Transformation by the Bmi-1 oncoprotein correlates with its subnuclear localization but not its transcriptional suppression activity.
The bmi-1 oncogene cooperates with c-myc in transgenic mice, resulting in accelerated lymphoma development. Altering the expression of Bmi-1 affects normal embryogenesis. The protein product of bmi-1 is homologous to certain Drosophila Polycomb group proteins that regulate homeotic gene expression through alteration of chromatin structure. Chimeric LexA-Bmi-1 protein has previously been shown to repress transcription. How Bmi-1 functions in embryogenesis and whether this relates to the ability of Bmi-1 to mediate cellular transformation is unknown. We demonstrate here that Bmi-1 is able to transform rodent fibroblasts in vitro, providing a system that has allowed us to correlate its molecular properties with its ability to transform cells. We map functional domains of Bmi-1 involved in transcriptional suppression by using the GAL4 chimeric transcriptional regulator system. Deletion analysis shows that the centrally located helix-turn-helix-turn-helix-turn (HTHTHT) motif is necessary for transcriptional suppression whereas the N-terminal RING finger domain is not required. We demonstrate that nuclear localization requires KRMK (residues 230 to 233) and that the absence of nuclear entry ablates transformation. In addition, we find that the subnuclear localization of wild-type Bmi-1 to the rim of the nucleus requires the RING finger domain and correlates with its ability to transform. Our studies with Bmi-1 deletion mutants suggest that the ability of Bmi-1 to mediate cellular transformation correlates with its unique subnuclear localization but not its transcriptional suppression activity.
Cohen
K J
KJ
Division of Pediatric Oncology, Johns Hopkins Oncology Center, Baltimore, Maryland 21205, USA.
Hanna
J S
JS
Prescott
J E
JE
Dang
C V
CV
D000818:Animals; D000906:Antibodies; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002471:Cell Transformation, Neoplastic; D003062:Codon; D004330:Drosophila; D029721:Drosophila Proteins; D005314:Embryonic and Fetal Development; D018507:Gene Expression Regulation, Developmental; D019077:Helix-Turn-Helix Motifs; D008223:Lymphoma / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D016366:Open Reading Frames; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D063150:Polycomb Repressive Complex 1; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000737:chemistry; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D012097:Repressor Proteins; D014158:Transcription, Genetic; D016335:Zinc Fingers
D000906:Antibodies; C489421:Bmi1 protein, mouse; C489420:Bmi1 protein, rat; D003062:Codon; D029721:Drosophila Proteins; D009687:Nuclear Proteins; C066770:Pc protein, Drosophila; D010446:Peptide Fragments; D011506:Proteins; D011518:Proto-Oncogene Proteins; D016271:Proto-Oncogene Proteins c-myc; D011994:Recombinant Proteins; D012097:Repressor Proteins; D063150:Polycomb Repressive Complex 1