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Sequence‐specific binding of a c‐myc nuclear‐matrix‐associated region shows increased nuclear matrix retention after leukemic cell (HL‐60) differentiation
Journal article

Sequence‐specific binding of a c‐myc nuclear‐matrix‐associated region shows increased nuclear matrix retention after leukemic cell (HL‐60) differentiation

Robin H. Chou, Judy Rae Churchill, Diane E. Mapstone and Marcella M. Flubacher
American journal of anatomy, v 191(3), pp 312-320
Jul 1991
PMID: 1927975

Abstract

HL‐60 cells, a human promyelocytic leukemia cell line, contain amplified c‐myc DNA sequences and mRNA transcripts. These cells can be induced to undergo macrophage differentiation by phorbol esters, which results in suppression of c‐myc expression and cessation of cell proliferation. The nuclear matrix (NM), a nuclear skeleton resistant to DNase I digestion and high salt extraction, is proposed to be involved in DNA replication, gene regulation, and the correct distribution of DNA at mitosis. We have previously identified a nuclear‐matrix‐associated region (MAR) of the c‐myc protooncogene to reside in a 1.4‐kb region between Cla I and Eco RI restriction sites at the 3′‐end of the gene. A 172‐bp Dra I/Dra I subfragment of the 1.4‐kb region was shown to be a major component of the MAR (myc‐MAE), and this subfragment was demonstrated to be recognized by a nuclear protein (p25). In this report we demonstrate that ϕX174 DNA, or the synthetic copolymers poly[d(G·C)] and poly[d(A·T)], are not effective suppressors of the binding of the myc‐MAR to isolated NM, indicating that the binding sequence(s) are unique. We find that the addition of partially purified protein p25 increases the relative affinity of the myc‐MAR for HL‐60 NM in an in vitro assay system. NM isolated from HL‐60 macrophages induced by phorbol esters retains significantly more myc‐MAR DNA fragment in the presence of an excess amount of competitor DNA than does NM from untreated HL‐60 cells. These data suggest that a change of the myc‐MAR association with the NM occurs after monocytic differentiation of HL‐60 cells.

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Anatomy & Morphology
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