title: PDIP38 is translocated to the spliceosomes/nuclear speckles in response to UV-induced DNA damage and is required for UV-induced alternative splicing of MDM2.
abstract: PDIP38 (polymerase delta interacting protein 38) was originally discovered as a protein that interacts with DNA polymerase δ and PCNA. PDIP38 is present in multiple intracellular locations and is a multifunctional protein that has been implicated in several diverse cellular functions. We investigated the nuclear localization of PDIP38 in order to gain insights to its response to UV damage. PDIP38 was found to form distinct nuclear foci in response to UV irradiation in several cell lines, including HeLa S3 and A549 cells. However, these foci were not those associated with UV repair foci. Using various markers for different nuclear subcompartments, the UV-induced PDIP38 foci were identified as spliceosomes/nuclear speckles, the storage and assembly sites for mRNA splicing factors. To assess the role of PDIP38 in the regulation of splicing events, the effects of PDIP38 depletion on the UV-induced alternate splicing of MDM2 transcripts were examined by nested RT-PCR. Alternatively spliced MDM2 products were induced by UV treatment but were greatly reduced in cells expressing shRNA targeting PDIP38. These findings indicate that upon UV-induced DNA damage, PDIP38 is translocated to spliceosomes and contributes to the UV-induced alternative splicing of MDM2 transcripts. Similar results were obtained when cells were subjected to transcriptional stresses with actinomycin D or α-amanitin. Taken together, these studies show that PDIP38 is a protein regulated in a dynamic manner in response to genotoxic stress, as evidenced by its translocation to the spliceosomes. Moreover, PDIP38 is required for the induction of the alternative splicing of MDM2 in response to UV irradiation.
authors.affiliation: Department of Biochemistry and Molecular Biology; New York Medical College; Valhalla, NY USA.
authors.forename: Agnes
authors.initials: A
authors.lastname: Wong
authors.forename: Sufang
authors.initials: S
authors.lastname: Zhang
authors.forename: Dana
authors.initials: D
authors.lastname: Mordue
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Wu
authors.forename: Zhongtao
authors.initials: Z
authors.lastname: Zhang
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Darzynkiewicz
authors.forename: Ernest Y C
authors.initials: EY
authors.lastname: Lee
authors.forename: Marietta Y W T
authors.initials: MY
authors.lastname: Lee
mesh_terms: D017398:Alternative Splicing; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects / Q000528:radiation effects*; D003609:Dactinomycin / Q000633:toxicity; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2 / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D017381:Spliceosomes / Q000378:metabolism*; D014466:Ultraviolet Rays
chemical_list: D009687:Nuclear Proteins; C473592:POLDIP2 protein, human; D018809:Proliferating Cell Nuclear Antigen; D034741:RNA, Small Interfering; D003609:Dactinomycin; D051736:Proto-Oncogene Proteins c-mdm2
keywords: DNA damage; DNA polymerase δ; MDM2; PDIP38; Poldip2; alternative splicing; nuclear speckles; spliceosomes