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Leukemia ; 14(11): 1898-908, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11069025

RESUMO

The mixed-lineage leukemia gene (MLL) is associated with more than 25 chromosomal translocations involving band 11q23 in diverse subtypes of human acute leukemia. Conditional expression of a 50 kDa amino terminal fragment spanning the AT hook motifs of MLL (MLL3AT) causes cell cycle arrest, upregulation of p21Cip1 and p27KiP1 and partial monocytic differentiation of the monoblastic U937 cell line, suggesting a major role for MLL3AT in MLL-AF9-induced myelomonocytic differentiation. In this study, we analyzed the subcellular localization of conditionally expressed MLL3AT in both U937 and HeLa cell lines. Immunofluorescence staining, confocal laser scanning microscopy and immunoelectron microscopy indicated that MLL3AT, like endogenous MLL, localized in the nucleoplasm in a punctate pattern of distribution, including regions attached to the nuclear envelope and the periphery of the nucleolus. We found that MLL3AT and endogenous MLL were present in interphase nuclear matrices and colocalized with topoisomerase II to mitotic chromosomal scaffolds. Nucleoplasm and nucleolar localization was observed even for MLL-AF9 and MLL-AF4 conditionally expressed chimeric proteins, suggesting a common target conferred by the amino terminus of MLL to many if not all the chimeric MLL proteins. The nuclear matrix/scaffold association suggests a role for the amino terminus of MLL in the modulation of chromatin structure, leading to epigenetic effects on the maintenance of gene expression.


Assuntos
Nucléolo Celular/metabolismo , Estruturas Cromossômicas/metabolismo , Proteínas de Ligação a DNA/metabolismo , Matriz Nuclear/metabolismo , Sinais Direcionadores de Proteínas , Proto-Oncogenes , Fatores de Transcrição , Transporte Ativo do Núcleo Celular , Motivos de Aminoácidos , Ciclo Celular , Proteínas Cromossômicas não Histona/metabolismo , DNA Topoisomerases Tipo II/análise , Proteínas de Ligação a DNA/fisiologia , Técnica Indireta de Fluorescência para Anticorpo , Células HeLa/metabolismo , Células HeLa/ultraestrutura , Histona-Lisina N-Metiltransferase , Humanos , Microscopia Confocal , Microscopia Imunoeletrônica , Mitose , Proteína de Leucina Linfoide-Mieloide , Proteínas de Neoplasias/metabolismo , Sinais de Localização Nuclear , Estrutura Terciária de Proteína , Transporte Proteico , Proteínas Recombinantes de Fusão/metabolismo , Transfecção , Células U937/metabolismo , Células U937/ultraestrutura
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