The Golgi complex is a regulatory hub for homologous recombination-mediated DNA repair

The Golgi complex has long been recognised as an important homeostasis hub, where a multitude of signalling pathways and essential cellular processes intersect. Yet its communication with the cell nucleus remains largely unexplored. To this end, we have analysed genome-scale localisation data of the Human Protein Atlas which revealed an unexpected high number of Golgi and nuclear dual-localisation proteins and several pathways including surprising DNA repair. Amongst these proteins we found RAD51C, a regulatory Homologous Recombination (HR) repair protein, that localises to the Golgi and in response to double-strand DNA breaks, the Golgi protein population of RAD51C redistributes to form DNA repair foci. Depletion of the Golgin Giantin induces the redistribution of the RAD51C Golgi pool to form nuclear foci, independent of DNA damage induction. Concurrent with a significant increase in genomic instability and inhibition of HR signalling regulators. Altogether, we present evidence for a novel pathway where the Golgi is a central regulatory hub for HR-mediated DNA repair and potentially other repair pathways.

[1]  Waldir Leoncio Netto,et al.  A combined experimental-computational approach uncovers a role for the Golgi matrix protein Giantin in breast cancer progression , 2022, bioRxiv.

[2]  Sarah Bui,et al.  Adaptation of the Golgi Apparatus in Cancer Cell Invasion and Metastasis , 2021, Frontiers in Cell and Developmental Biology.

[3]  Robert A. Baldock,et al.  RAD51 paralog function in replicative DNA damage and tolerance. , 2021, Current opinion in genetics & development.

[4]  Xiaoyan Zhang.  Alterations of Golgi Structural Proteins and Glycosylation Defects in Cancer , 2021, Frontiers in Cell and Developmental Biology.

[5]  Zhiping Hu,et al.  The role of the Golgi apparatus in disease (Review) , 2021, International journal of molecular medicine.

[6]  Nadezhda T. Doncheva,et al.  The STRING database in 2021: customizable protein–protein networks, and functional characterization of user-uploaded gene/measurement sets , 2020, Nucleic Acids Res..

[7]  P. Møller,et al.  An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity , 2020, Nature Protocols.

[8]  G. Legube,et al.  Studying DNA Double-Strand Break Repair: An Ever-Growing Toolbox , 2020, Frontiers in Molecular Biosciences.

[9]  Prajakta Kulkarni-Gosavi,et al.  Form and function of the Golgi apparatus: scaffolds, cytoskeleton and signalling , 2019, FEBS letters.

[10]  Kara A. Bernstein,et al.  RAD-ical New Insights into RAD51 Regulation , 2018, Genes.

[11]  Christian Makhoul,et al.  The Golgi architecture and cell sensing. , 2018, Biochemical Society transactions.

[12]  J. Goldstein,et al.  Retrospective on Cholesterol Homeostasis: The Central Role of Scap. , 2018, Annual review of biochemistry.

[13]  P. Møller.  The comet assay: ready for 30 more years. , 2018, Mutagenesis.

[14]  M. D. De Matteis,et al.  The Golgi complex in disease and therapy. , 2018, Current opinion in cell biology.

[15]  P. Cheng,et al.  Shifted Golgi targeting of glycosyltransferases and α-mannosidase IA from giantin to GM130-GRASP65 results in formation of high mannose N-glycans in aggressive prostate cancer cells. , 2017, Biochimica et biophysica acta. General subjects.

[16]  Devin P. Sullivan,et al.  A subcellular map of the human proteome , 2017, Science.

[17]  P. Bork,et al.  The endosomal transcriptional regulator RNF11 integrates degradation and transport of EGFR , 2016, The Journal of cell biology.

[18]  M. Lowe,et al.  The Golgin Family of Coiled-Coil Tethering Proteins , 2016, Front. Cell Dev. Biol..

[19]  C. Machamer.  The Golgi complex in stress and death , 2015, Front. Neurosci..

[20]  H. Farhan,et al.  Loss of GM130 in breast cancer cells and its effects on cell migration, invasion and polarity , 2015, Cell cycle.

[21]  M. Jasin,et al.  Homologous recombination and human health: the roles of BRCA1, BRCA2, and associated proteins. , 2015, Cold Spring Harbor perspectives in biology.

[22]  A. Petrosyan.  Onco-Golgi: Is Fragmentation a Gate to Cancer Progression? , 2015, Biochemistry & molecular biology journal.

[23]  K. O'Byrne,et al.  Chemotherapeutic Compounds Targeting the DNA Double-Strand Break Repair Pathways: The Good, the Bad, and the Promising , 2014, Front. Oncol..

[24]  Michelle M. Ng,et al.  DNA Damage Triggers Golgi Dispersal via DNA-PK and GOLPH3 , 2014, Cell.

[25]  David Hsu,et al.  OpenComet: An automated tool for comet assay image analysis , 2014, Redox biology.

[26]  Tang Zi.  Pathway choice for DNA double-strand break repair , 2014 .

[27]  A. Colanzi,et al.  Signaling at the Golgi during mitosis. , 2013, Methods in cell biology.

[28]  B. Cornelissen,et al.  Targeting ATR in vivo using the novel inhibitor VE-822 results in selective sensitization of pancreatic tumors to radiation , 2012, Cell Death and Disease.

[29]  D. Gent,et al.  Pathway choice in DNA double strand break repair: observations of a balancing act , 2012, Genome Integrity.

[30]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[31]  Martin Hjelmare,et al.  Systematic validation of antibody binding and protein subcellular localization using siRNA and confocal microscopy. , 2012, Journal of proteomics.

[32]  P. Olive,et al.  Heterogeneity in Radiation-Induced DNA Damage and Repair in Tumor and Normal Cells Measured Using the “Comet” Assay , 1990 .

[33]  Susan L. Neuhausen,et al.  RAD51C Germline Mutations in Breast and Ovarian Cancer Cases from High-Risk Families , 2011, PloS one.

[34]  H. Freeze,et al.  Golgi glycosylation and human inherited diseases. , 2011, Cold Spring Harbor perspectives in biology.

[35]  Tina Hernandez-Boussard,et al.  Doxorubicin pathways: pharmacodynamics and adverse effects , 2011, Pharmacogenetics and genomics.

[36]  Karsten Weis,et al.  Importazole, a small molecule inhibitor of the transport receptor importin-β. , 2011, ACS chemical biology.

[37]  N. Curtin,et al.  Further characterisation of the cellular activity of the DNA-PK inhibitor, NU7441, reveals potential cross-talk with homologous recombination , 2011, Cancer Chemotherapy and Pharmacology.

[38]  A. Colanzi,et al.  The Golgi apparatus: an organelle with multiple complex functions. , 2011, The Biochemical journal.

[39]  Kumar Somyajit,et al.  RAD51C: a novel cancer susceptibility gene is linked to Fanconi anemia and breast cancer , 2010, Carcinogenesis.

[40]  Franca Fraternali,et al.  Mutation of the RAD51C gene in a Fanconi anemia–like disorder , 2010, Nature Genetics.

[41]  Jay M. Sage,et al.  Cellular Redistribution of Rad51 in Response to DNA Damage , 2009, The Journal of Biological Chemistry.

[42]  R. Wollman,et al.  A genome-wide siRNA screen reveals diverse cellular processes and pathways that mediate genome stability. , 2009, Molecular cell.

[43]  C. Liao,et al.  RAD51C facilitates checkpoint signaling by promoting CHK2 phosphorylation , 2009, The Journal of cell biology.

[44]  C. L. Jackson,et al.  Mechanisms of transport through the Golgi complex , 2009, Journal of Cell Science.

[45]  Holger Erfle,et al.  Work Flow for Multiplexing siRNA Assays by Solid-Phase Reverse Transfection in Multiwell Plates , 2008, Journal of biomolecular screening.

[46]  Anne E Carpenter,et al.  CellProfiler: image analysis software for identifying and quantifying cell phenotypes , 2006, Genome Biology.

[47]  N. Curtin,et al.  Identification and Characterization of a Novel and Specific Inhibitor of the Ataxia-Telangiectasia Mutated Kinase ATM , 2004, Cancer Research.

[48]  C. A. French,et al.  Identification of Functional Domains in the RAD51L2 (RAD51C) Protein and Its Requirement for Gene Conversion* , 2003, Journal of Biological Chemistry.

[49]  K. Mori,et al.  Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress. , 1999, Molecular biology of the cell.