title: DICER, DROSHA and DNA damage response RNAs are necessary for the secondary recruitment of DNA damage response factors.
abstract: The DNA damage response (DDR) plays a central role in preserving genome integrity. Recently, we reported that the endoribonucleases DICER and DROSHA contribute to DDR activation by generating small non-coding RNAs, termed DNA damage response RNA (DDRNA), carrying the sequence of the damaged locus. It is presently unclear whether DDRNAs act by promoting the primary recognition of DNA lesions or the secondary recruitment of DDR factors into cytologically detectable foci and consequent signal amplification. Here, we demonstrate that DICER and DROSHA are dispensable for primary recruitment of the DDR sensor NBS1 to DNA damage sites. Instead, the accumulation of the DDR mediators MDC1 and 53BP1 (also known as TP53BP1), markers of secondary recruitment, is reduced in DICER- or DROSHA-inactivated cells. In addition, NBS1 (also known as NBN) primary recruitment is resistant to RNA degradation, consistent with the notion that RNA is dispensable for primary recognition of DNA lesions. We propose that DICER, DROSHA and DDRNAs act in the response to DNA damage after primary recognition of DNA lesions and, together with γH2AX, are essential for enabling the secondary recruitment of DDR factors and fuel the amplification of DDR signaling.
authors.affiliation: IFOM Foundation - The FIRC Institute of Molecular Oncology Foundation, Via Adamello 16, Milan 20139, Italy Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche (IGM-CNR), Via Abbiategrasso 207, Pavia 27100, Italy sofia.francia@ifom.eu fabrizio.dadda@ifom.eu.
authors.forename: Sofia
authors.initials: S
authors.lastname: Francia
authors.affiliation: Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche (IGM-CNR), Via Abbiategrasso 207, Pavia 27100, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Cabrini
authors.affiliation: IFOM Foundation - The FIRC Institute of Molecular Oncology Foundation, Via Adamello 16, Milan 20139, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Matti
authors.affiliation: IFOM Foundation - The FIRC Institute of Molecular Oncology Foundation, Via Adamello 16, Milan 20139, Italy.
authors.forename: Amanda
authors.initials: A
authors.lastname: Oldani
authors.affiliation: IFOM Foundation - The FIRC Institute of Molecular Oncology Foundation, Via Adamello 16, Milan 20139, Italy Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche (IGM-CNR), Via Abbiategrasso 207, Pavia 27100, Italy sofia.francia@ifom.eu fabrizio.dadda@ifom.eu.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: d'Adda di Fagagna
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D053487:DEAD-box RNA Helicases / Q000235:genetics*; D004249:DNA Damage / Q000235:genetics*; D004260:DNA Repair / Q000235:genetics / Q000276:immunology*; D006657:Histones / Q000378:metabolism*; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D043244:Ribonuclease III / Q000235:genetics*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D000071857:Tumor Suppressor p53-Binding Protein 1 / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; C468783:H2AX protein, human; D006657:Histones; C413163:MDC1 protein, human; C112156:NBN protein, human; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; C498301:TP53BP1 protein, human; D015534:Trans-Activators; D000071857:Tumor Suppressor p53-Binding Protein 1; C408087:DICER1 protein, human; C517918:DROSHA protein, human; D043244:Ribonuclease III; D012259:Ribonuclease, Pancreatic; D053487:DEAD-box RNA Helicases
keywords: DICER; DNA damage response; DROSHA; Non-coding RNA