Chromatin meets the cytoskeleton: the importance of nuclear actin dynamics and associated motors for genome stability
DNA Repair (Amst). 2023 Sep 16;131:103571. doi: 10.1016/j.dnarep.2023.103571. Online ahead of print.ABSTRACTThe actin cytoskeleton is of fundamental importance for numerous cellular processes, including intracellular transport, cell plasticity, and cell migration. However, functions of filamentous actin (F-actin) in the nucleus remain understudied due to the comparatively low abundance of nuclear actin and the resulting experimental limitations to its visualization. Owing to recent technological advances such as super-resolution microscopy and the development of nuclear-specific actin probes, essential roles of the actin c...
Source: DNA Repair - September 22, 2023 Category: Genetics & Stem Cells Authors: Hans-Peter Wollscheid Helle D Ulrich Source Type: research

Chromatin meets the cytoskeleton: the importance of nuclear actin dynamics and associated motors for genome stability
DNA Repair (Amst). 2023 Sep 16;131:103571. doi: 10.1016/j.dnarep.2023.103571. Online ahead of print.ABSTRACTThe actin cytoskeleton is of fundamental importance for numerous cellular processes, including intracellular transport, cell plasticity, and cell migration. However, functions of filamentous actin (F-actin) in the nucleus remain understudied due to the comparatively low abundance of nuclear actin and the resulting experimental limitations to its visualization. Owing to recent technological advances such as super-resolution microscopy and the development of nuclear-specific actin probes, essential roles of the actin c...
Source: DNA Repair - September 22, 2023 Category: Genetics & Stem Cells Authors: Hans-Peter Wollscheid Helle D Ulrich Source Type: research

Multi-scale cellular imaging of DNA double strand break repair
We describe different (live-cell) imaging modalities that can reveal details of the DNA double-strand break repair process across multiple time and spatial scales. In addition, recent developments are discussed in super-resolution imaging and single-molecule tracking, and how these technologies can be applied to elucidate details on structural compositions or dynamics of DNA double-strand break repair.PMID:37734176 | DOI:10.1016/j.dnarep.2023.103570 (Source: DNA Repair)
Source: DNA Repair - September 21, 2023 Category: Genetics & Stem Cells Authors: Tim Heemskerk Gerarda van de Kamp Jeroen Essers Roland Kanaar Maarten W Paul Source Type: research

Multi-scale cellular imaging of DNA double strand break repair
We describe different (live-cell) imaging modalities that can reveal details of the DNA double-strand break repair process across multiple time and spatial scales. In addition, recent developments are discussed in super-resolution imaging and single-molecule tracking, and how these technologies can be applied to elucidate details on structural compositions or dynamics of DNA double-strand break repair.PMID:37734176 | DOI:10.1016/j.dnarep.2023.103570 (Source: DNA Repair)
Source: DNA Repair - September 21, 2023 Category: Genetics & Stem Cells Authors: Tim Heemskerk Gerarda van de Kamp Jeroen Essers Roland Kanaar Maarten W Paul Source Type: research

Live cell transcription-coupled nucleotide excision repair dynamics revisited
In this study, we employ knock-in cell lines designed to target the endogenous CSB gene locus with mClover, a GFP variant. Through live cell imaging, we uncover the intricate molecular dynamics of CSB in response to physiologically relevant UV doses. We showed that the DNA damage-induced association of CSB with chromatin is tightly regulated by the CSA-containing ubiquitin-ligase CRL complex (CRL4CSA). Combining the CSB-mClover knock-in cell line with SILAC-based GFP-mediated complex isolation and mass-spectrometry-based proteomics, revealed novel putative CSB interactors as well as discernible variations in complex compos...
Source: DNA Repair - September 16, 2023 Category: Genetics & Stem Cells Authors: Diana A Llerena Schiffmacher Katarzyna W Kliza Arjan F Theil Gert-Jan Kremers Jeroen A A Demmers Tomoo Ogi Michiel Vermeulen Wim Vermeulen Alex Pines Source Type: research

Live cell transcription-coupled nucleotide excision repair dynamics revisited
In this study, we employ knock-in cell lines designed to target the endogenous CSB gene locus with mClover, a GFP variant. Through live cell imaging, we uncover the intricate molecular dynamics of CSB in response to physiologically relevant UV doses. We showed that the DNA damage-induced association of CSB with chromatin is tightly regulated by the CSA-containing ubiquitin-ligase CRL complex (CRL4CSA). Combining the CSB-mClover knock-in cell line with SILAC-based GFP-mediated complex isolation and mass-spectrometry-based proteomics, revealed novel putative CSB interactors as well as discernible variations in complex compos...
Source: DNA Repair - September 16, 2023 Category: Genetics & Stem Cells Authors: Diana A Llerena Schiffmacher Katarzyna W Kliza Arjan F Theil Gert-Jan Kremers Jeroen A A Demmers Tomoo Ogi Michiel Vermeulen Wim Vermeulen Alex Pines Source Type: research

Live cell transcription-coupled nucleotide excision repair dynamics revisited
In this study, we employ knock-in cell lines designed to target the endogenous CSB gene locus with mClover, a GFP variant. Through live cell imaging, we uncover the intricate molecular dynamics of CSB in response to physiologically relevant UV doses. We showed that the DNA damage-induced association of CSB with chromatin is tightly regulated by the CSA-containing ubiquitin-ligase CRL complex (CRL4CSA). Combining the CSB-mClover knock-in cell line with SILAC-based GFP-mediated complex isolation and mass-spectrometry-based proteomics, revealed novel putative CSB interactors as well as discernible variations in complex compos...
Source: DNA Repair - September 16, 2023 Category: Genetics & Stem Cells Authors: Diana A Llerena Schiffmacher Katarzyna W Kliza Arjan F Theil Gert-Jan Kremers Jeroen A A Demmers Tomoo Ogi Michiel Vermeulen Wim Vermeulen Alex Pines Source Type: research

Live cell transcription-coupled nucleotide excision repair dynamics revisited
In this study, we employ knock-in cell lines designed to target the endogenous CSB gene locus with mClover, a GFP variant. Through live cell imaging, we uncover the intricate molecular dynamics of CSB in response to physiologically relevant UV doses. We showed that the DNA damage-induced association of CSB with chromatin is tightly regulated by the CSA-containing ubiquitin-ligase CRL complex (CRL4CSA). Combining the CSB-mClover knock-in cell line with SILAC-based GFP-mediated complex isolation and mass-spectrometry-based proteomics, revealed novel putative CSB interactors as well as discernible variations in complex compos...
Source: DNA Repair - September 16, 2023 Category: Genetics & Stem Cells Authors: Diana A Llerena Schiffmacher Katarzyna W Kliza Arjan F Theil Gert-Jan Kremers Jeroen A A Demmers Tomoo Ogi Michiel Vermeulen Wim Vermeulen Alex Pines Source Type: research

The C-terminal tail of Rad17, iVERGE, binds the 9 ‒1‒1 complex independently of AAA+ ATPase domains to provide another clamploader interface
DNA Repair (Amst). 2023 Sep 6;130:103567. doi: 10.1016/j.dnarep.2023.103567. Online ahead of print.ABSTRACTThe ATR pathway plays a crucial role in maintaining genome integrity as the major DNA damage checkpoint. It also attracts attention as a therapeutic target in cancer treatment. The Rad17-RFC2-5 complex loads the Rad9-Hus1-Rad1 (9-1-1) DNA clamp complex onto damaged chromatin to activate the ATR pathway. We previously reported that phosphorylation of a polyanionic C-terminal tail of human Rad17, iVERGE, is essential for the interaction between Rad17 and the 9-1-1 complex. However, the molecular mechanism has remained u...
Source: DNA Repair - September 15, 2023 Category: Genetics & Stem Cells Authors: Yasunori Fukumoto Tyuji Hoshino Yuji Nakayama Yasumitsu Ogra Source Type: research

The C-terminal tail of Rad17, iVERGE, binds the 9 ‒1‒1 complex independently of AAA+ ATPase domains to provide another clamploader interface
DNA Repair (Amst). 2023 Sep 6;130:103567. doi: 10.1016/j.dnarep.2023.103567. Online ahead of print.ABSTRACTThe ATR pathway plays a crucial role in maintaining genome integrity as the major DNA damage checkpoint. It also attracts attention as a therapeutic target in cancer treatment. The Rad17-RFC2-5 complex loads the Rad9-Hus1-Rad1 (9-1-1) DNA clamp complex onto damaged chromatin to activate the ATR pathway. We previously reported that phosphorylation of a polyanionic C-terminal tail of human Rad17, iVERGE, is essential for the interaction between Rad17 and the 9-1-1 complex. However, the molecular mechanism has remained u...
Source: DNA Repair - September 15, 2023 Category: Genetics & Stem Cells Authors: Yasunori Fukumoto Tyuji Hoshino Yuji Nakayama Yasumitsu Ogra Source Type: research

Centromere: A Trojan horse for genome stability
DNA Repair (Amst). 2023 Sep 7;130:103569. doi: 10.1016/j.dnarep.2023.103569. Online ahead of print.ABSTRACTCentromeres play a key role in the maintenance of genome stability to prevent carcinogenesis and diseases. They are specialized chromosome loci essential to ensure faithful transmission of genomic information across cell generations by mediating the interaction with spindle microtubules. Nonetheless, while fulfilling these essential roles, their distinct repetitive composition and susceptibility to mechanical stresses during cell division render them susceptible to breakage events. In this review, we delve into the pr...
Source: DNA Repair - September 14, 2023 Category: Genetics & Stem Cells Authors: Andrea Scelfo Daniele Fachinetti Source Type: research

Centromere: A Trojan horse for genome stability
DNA Repair (Amst). 2023 Sep 7;130:103569. doi: 10.1016/j.dnarep.2023.103569. Online ahead of print.ABSTRACTCentromeres play a key role in the maintenance of genome stability to prevent carcinogenesis and diseases. They are specialized chromosome loci essential to ensure faithful transmission of genomic information across cell generations by mediating the interaction with spindle microtubules. Nonetheless, while fulfilling these essential roles, their distinct repetitive composition and susceptibility to mechanical stresses during cell division render them susceptible to breakage events. In this review, we delve into the pr...
Source: DNA Repair - September 14, 2023 Category: Genetics & Stem Cells Authors: Andrea Scelfo Daniele Fachinetti Source Type: research

Metazoan nuclear pore complexes in gene regulation and genome stability
DNA Repair (Amst). 2023 Sep 4;130:103565. doi: 10.1016/j.dnarep.2023.103565. Online ahead of print.ABSTRACTThe nuclear pore complexes (NPCs), one of the hallmarks of eukaryotic nuclei, allow selective transport of macromolecules between the cytoplasm and the nucleus. Besides this canonical function, an increasing number of additional roles have been attributed to the NPCs and their constituents, the nucleoporins. Here we review recent insights into the mechanisms by which NPCs and nucleoporins affect transcription and DNA repair in metazoans. In the first part, we discuss how gene expression can be affected by the localiza...
Source: DNA Repair - September 11, 2023 Category: Genetics & Stem Cells Authors: Parisa Nobari Val érie Doye Charlene Boumendil Source Type: research

Metazoan nuclear pore complexes in gene regulation and genome stability
DNA Repair (Amst). 2023 Sep 4;130:103565. doi: 10.1016/j.dnarep.2023.103565. Online ahead of print.ABSTRACTThe nuclear pore complexes (NPCs), one of the hallmarks of eukaryotic nuclei, allow selective transport of macromolecules between the cytoplasm and the nucleus. Besides this canonical function, an increasing number of additional roles have been attributed to the NPCs and their constituents, the nucleoporins. Here we review recent insights into the mechanisms by which NPCs and nucleoporins affect transcription and DNA repair in metazoans. In the first part, we discuss how gene expression can be affected by the localiza...
Source: DNA Repair - September 11, 2023 Category: Genetics & Stem Cells Authors: Parisa Nobari Val érie Doye Charlene Boumendil Source Type: research

Metazoan nuclear pore complexes in gene regulation and genome stability
DNA Repair (Amst). 2023 Sep 4;130:103565. doi: 10.1016/j.dnarep.2023.103565. Online ahead of print.ABSTRACTThe nuclear pore complexes (NPCs), one of the hallmarks of eukaryotic nuclei, allow selective transport of macromolecules between the cytoplasm and the nucleus. Besides this canonical function, an increasing number of additional roles have been attributed to the NPCs and their constituents, the nucleoporins. Here we review recent insights into the mechanisms by which NPCs and nucleoporins affect transcription and DNA repair in metazoans. In the first part, we discuss how gene expression can be affected by the localiza...
Source: DNA Repair - September 11, 2023 Category: Genetics & Stem Cells Authors: Parisa Nobari Val érie Doye Charlene Boumendil Source Type: research