Functional proteomic analysis of promyelocytic leukaemia nuclear bodies in irradiation-induced MCF-7 cells.
It is well established that promyelocytic leukaemia nuclear bodies (PML NBs) play important roles in DNA damage responses (DDR). After irradiation, PML NBs dynamically recruit or release important proteins involved in cell-cycle regulation, DNA repair and apoptosis. As PML protein is the key molecule of PML NBs' dynamic assembling, we aimed to characterize the PML-interacting proteins in (60)Co-irradiated MCF-7 cells. A proteomic approach using CoIP, mono-dimensional electrophoresis and tandem mass spectrometry, allowed us to identify a total of 124 proteins that may associate with PML after irradiation. Bioinformatic analysis of the identified proteins showed that most of them were related to characterized PML functions, such as transcriptional regulation, cell-cycle regulation, cell-death regulation and response to stress. Four proteins, B23, MVP, G3BP1 and DHX9, were verified to co-localize with PML differentially before and after ionizing radiation (IR) treatment. The proteins identified in this study will significantly improve our understanding of the dynamic organization and multiple functions of PML NBs in DDR.
Liu
Jinfeng
J
Department of Biochemistry and Molecular Biology, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, China.
Song
Yi
Y
Tian
Baolei
B
Qian
Junjie
J
Dong
Yan
Y
Liu
Jilai
J
Liu
Bin
B
Sun
Zhixian
Z
D017209:Apoptosis / Q000528:radiation effects*; D001943:Breast Neoplasms / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000502:physiology; D045744:Cell Line, Tumor; D022003:Cell Nucleus Structures / Q000378:metabolism / Q000528:radiation effects / Q000648:ultrastructure; D004249:DNA Damage / Q000528:radiation effects*; D004260:DNA Repair / Q000528:radiation effects*; D004273:DNA, Neoplasm / Q000378:metabolism*; D005260:Female; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000378:metabolism* / Q000648:ultrastructure; D009687:Nuclear Proteins / Q000378:metabolism / Q000648:ultrastructure; D000071156:Promyelocytic Leukemia Protein; D040901:Proteomics; D014157:Transcription Factors / Q000378:metabolism / Q000648:ultrastructure; D025521:Tumor Suppressor Proteins / Q000378:metabolism / Q000502:physiology / Q000528:radiation effects* / Q000648:ultrastructure
D018797:Cell Cycle Proteins; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human