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© The Rockefeller University Press, 0021-9525/2001//367 $5.00
The Journal of Cell Biology, Volume 153, Number 2, , 2001 367-380

Regulation and Localization of the Bloom Syndrome Protein in Response to DNA Damage

^a Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720
^b Berlex Laboratories, Inc., Richmond, California 94804
^c Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, New York 10021
Lawrence Berkeley National Laboratory, Mailstop 84-171, Berkeley, CA 94720.(510) 486-4545(510) 486-4416

jcampisi{at}lbl.gov

Bloom syndrome (BS) is an autosomal recessive disorder characterized by a high incidence of cancer and genomic instability. BLM, the protein defective in BS, is a RecQ-like helicase, presumed to function in DNA replication, recombination, or repair. BLM localizes to promyelocytic leukemia protein (PML) nuclear bodies and is expressed during late S and G2. We show, in normal human cells, that the recombination/repair proteins hRAD51 and replication protein (RP)-A assembled with BLM into a fraction of PML bodies during late S/G2. Biochemical experiments suggested that BLM resides in a nuclear matrix–bound complex in which association with hRAD51 may be direct. DNA-damaging agents that cause double strand breaks and a G2 delay induced BLM by a p53- and ataxia-telangiectasia mutated independent mechanism. This induction depended on the G2 delay, because it failed to occur when G2 was prevented or bypassed. It coincided with the appearance of foci containing BLM, PML, hRAD51 and RP-A, which resembled ionizing radiation-induced foci. After radiation, foci containing BLM and PML formed at sites of single-stranded DNA and presumptive repair in normal cells, but not in cells with defective PML. Our findings suggest that BLM is part of a dynamic nuclear matrix–based complex that requires PML and functions during G2 in undamaged cells and recombinational repair after DNA damage.

Key Words: RECQ helicases • p53 • ATM • nuclear matrix • homologous recombination

© 2001 The Rockefeller University Press

Abbreviations used in this paper: AT, ataxia telangiectasia; ATM, AT mutated; BS, Bloom syndrome; DSB, double-strand break; DNA-PK, DNA-dependent protein kinase; GST, glutathione S-transferase; HR, homologous recombination; HU, hydroxyurea; IR, ionizing radiation; IRIF, IR-induced foci; LN, labeled nuclei; NB, nuclear body; PARP, poly-ADP ribose polymerase; PML, promyelocytic leukemia protein; RP, replication protein; RT, reverse transcriptase; RTS, Rothmund-Thomson syndrome; SCE, sister chromatid exchange; WS, Werner syndrome.

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