scholarly journals UvrD helicase–RNA polymerase interactions are governed by UvrD’s carboxy-terminal Tudor domain

2020 ◽  
Author(s):  
Ashish A. Kawale ◽  
Björn M. Burmann
Keyword(s):  
Dna Repair ◽  
Rna Polymerase ◽  
Uv Light ◽  
Genome Damage ◽  
Functional Aspects ◽  
Crucial Component ◽  
Tudor Domain ◽  
Carboxy Terminal Region ◽  
Living Organisms ◽  
Carboxy Terminal

ABSTRACTAll living organisms have to cope with the constant threat of genome damage by UV light and other toxic reagents. To maintain the integrity of their genomes, organisms developed a variety of DNA repair pathways. One of these, the Transcription Coupled DNA-Repair (TCR) pathway, is triggered by stalled RNA Polymerase (RNAP) complexes at DNA damage sites on actively transcribed genes. A recently elucidated bacterial TCR pathway employs the UvrD helicase pulling back stalled RNAP complexes from the damage, stimulating recruitment of the DNA-repair machinery. However, structural and functional aspects of UvrD’s interaction with RNA Polymerase remain elusive. Here we used advanced solution NMR spectroscopy to investigate UvrD’s role within the TCR, identifying that the carboxy-terminal region of the UvrD helicase facilitates RNAP interactions by adopting a Tudor-domain like fold. Subsequently, we functionally analyzed this domain, identifying it as a crucial component for the UvrD–RNAP interaction besides having nucleic-acid affinity.

scholarly journals UvrD helicase–RNA polymerase interactions are governed by UvrD’s carboxy-terminal Tudor domain

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Ashish A. Kawale ◽  
Björn M. Burmann
Keyword(s):  
Dna Repair ◽  
Rna Polymerase ◽  
Uv Light ◽  
Genome Damage ◽  
Functional Aspects ◽  
Crucial Component ◽  
Tudor Domain ◽  
Carboxy Terminal Region ◽  
Living Organisms ◽  
Carboxy Terminal

AbstractAll living organisms have to cope with the constant threat of genome damage by UV light and other toxic reagents. To maintain the integrity of their genomes, organisms developed a variety of DNA repair pathways. One of these, the Transcription Coupled DNA-Repair (TCR) pathway, is triggered by stalled RNA Polymerase (RNAP) complexes at DNA damage sites on actively transcribed genes. A recently elucidated bacterial TCR pathway employs the UvrD helicase pulling back stalled RNAP complexes from the damage, stimulating recruitment of the DNA-repair machinery. However, structural and functional aspects of UvrD’s interaction with RNA Polymerase remain elusive. Here we used advanced solution NMR spectroscopy to investigate UvrD’s role within the TCR, identifying that the carboxy-terminal region of the UvrD helicase facilitates RNAP interactions by adopting a Tudor-domain like fold. Subsequently, we functionally analyzed this domain, identifying it as a crucial component for the UvrD–RNAP interaction besides having nucleic-acid affinity.

scholarly journals Publisher Correction: UvrD helicase–RNA polymerase interactions are governed by UvrD’s carboxy-terminal Tudor domain

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Ashish A. Kawale ◽  
Björn M. Burmann
Keyword(s):  
Rna Polymerase ◽  
Tudor Domain ◽  
Carboxy Terminal

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

scholarly journals Centrin/Cdc31 Is a Novel Regulator of Protein Degradation

2007 ◽  
Vol 28 (5) ◽  
pp. 1829-1840 ◽  
Author(s):  
Li Chen ◽  
Kiran Madura
Keyword(s):  
Dna Repair ◽  
Protein Degradation ◽  
Calcium Binding ◽  
Excision Repair ◽  
Uv Light ◽  
Dual Function ◽  
Highly Sensitive ◽  
Ef Hand ◽  
Carboxy Terminal ◽  
First Time

ABSTRACT Rad23 is required for efficient protein degradation and performs an important role in nucleotide excision repair. Saccharomyces cerevisiae Rad23, and its human counterpart (hHR23), are present in a complex containing the DNA repair factor Rad4 (termed XPC, for xeroderma pigmentosum group C, in humans). XPC/hHR23 was also reported to bind centrin-2, a member of the superfamily of calcium-binding EF-hand proteins. We report here that yeast centrin, which is encoded by CDC31, is similarly present in a complex with Rad4/Rad23 (called NEF2). The interaction between Cdc31 and Rad23/Rad4 varied by growth phase and reflected oscillations in Cdc31 levels. Strikingly, a cdc31 mutant that formed a weaker interaction with Rad4 showed sensitivity to UV light. Based on the dual function of Rad23, in both DNA repair and protein degradation, we questioned if Cdc31 also participated in protein degradation. We report here that Cdc31 binds the proteasome and multiubiquitinated proteins through its carboxy-terminal EF-hand motifs. Moreover, cdc31 mutants were highly sensitive to drugs that cause protein damage, failed to efficiently degrade proteolytic substrates, and formed altered interactions with the proteasome. These findings reveal for the first time a new role for centrin/Cdc31 in protein degradation.

scholarly journals Human HMGN1 and HMGN2 are not required for transcription-coupled DNA repair

2019 ◽  
Author(s):  
Katja Apelt ◽  
Iris Zoutendijk ◽  
Dennis Y. Gout ◽  
Diana van den Heuvel ◽  
Martijn S. Luijsterburg
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Uv Light ◽  
Dna Lesions ◽  
Illudin S ◽  
Highly Sensitive ◽  
Nucleosome Binding ◽  
Active Genes

SummaryTranscription-coupled repair (TCR) removes DNA lesions from the transcribed strand of active genes. Stalling of RNA polymerase II (RNAPII) at DNA lesions initiates TCR through the recruitment of the CSB and CSA proteins. The full repertoire of proteins required for human TCR – particularly in a chromatin context - remains to be determined. Studies in mice have revealed that the nucleosome-binding protein HMGN1 is required to enhance the repair of UV-induced lesions in transcribed genes. However, whether HMGN1 is required for human TCR remains unaddressed. Here, we show that knockout or knockdown of HMGN1, either alone or in combination with HMGN2, does not render human cells sensitive to UV light or Illudin S-induced transcription-blocking DNA lesions. Moreover, transcription restart after UV irradiation was not impaired in HMGN-deficient cells. In contrast, TCR-deficient cells were highly sensitive to DNA damage and failed to restart transcription. Furthermore, GFP-tagged HMGN1 was not recruited to sites of UV-induced DNA damage under conditions were GFP-CSB readily accumulated. In line with this, HMGN1 did not associate with the TCR complex, nor did TCR proteins require HMGN1 to associate with DNA damage-stalled RNAPII. Together, our findings suggest that HMGN1 and HMGN2 are not required for human TCR.

scholarly journals Formation and Recognition of UV-Induced DNA Damage within Genome Complexity

2020 ◽  
Vol 21 (18) ◽  
pp. 6689
Author(s):  
Philippe Johann to Berens ◽  
Jean Molinier
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Uv Light ◽  
Dna Repair Mechanisms ◽  
Genome Complexity ◽  
Repair Mechanisms ◽  
Living Organisms ◽  
Genomic Regions ◽  
The Impact

Ultraviolet (UV) light is a natural genotoxic agent leading to the formation of photolesions endangering the genomic integrity and thereby the survival of living organisms. To prevent the mutagenetic effect of UV, several specific DNA repair mechanisms are mobilized to accurately maintain genome integrity at photodamaged sites within the complexity of genome structures. However, a fundamental gap remains to be filled in the identification and characterization of factors at the nexus of UV-induced DNA damage, DNA repair, and epigenetics. This review brings together the impact of the epigenomic context on the susceptibility of genomic regions to form photodamage and focuses on the mechanisms of photolesions recognition through the different DNA repair pathways.

RECONSTRUCTION OF DNA-REPAIR DEFICIENT XERODERMA PIGMENTOSUM SKIN IN VITRO: A MODEL TO STUDY HYPERSENSITIVITY TO UV LIGHT

2004 ◽  
Author(s):  
Françoise Bernerd ◽  
Daniel Asselineau ◽  
Mathilde Frechet ◽  
Alain Sarasin ◽  
Thierry Magnaldo
Keyword(s):  
Dna Repair ◽  
Xeroderma Pigmentosum ◽  
Uv Light
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