An SR protein is essential for the recovery of malaria parasites from DNA damage and exposure to artemisinin

AbstractPlasmodium falciparum, the parasite responsible for the deadliest form of human malaria, maintains a complex life cycle with a relatively small number of genes. PfSR1 is an alternative splicing factor that regulates expansion of the P. falciparum protein repertoire. To further investigate PfSR1 functions, we set to unveil its interactome. We found that PfSR1 interacts with proteins, which are linked to various processes of RNA metabolism in a stage-dependent manner. These include: chromatin re-modeling, transcription, splicing and translation. Intriguingly, some of the PfSR1 interacting proteins are orthologues of proteins implicated in the DNA damage response. We demonstrate that PfSR1 expression is important for preventing the accumulation of DNA damage in proliferating parasites. In addition, following parasites’ exposure to a source of DNA damage, PfSR1 is recruited to damaged foci where it interacts with the phosphorylated core histone PfH2A, which marks damaged chromatin. Furthermore, PfSR1 expression was found to be essential for the ability of the parasite to activate the DNA repair machinery and recover from DNA damage caused by either irradiation or exposure to artemisinin, the first line anti-malarial drug. These findings unveil a novel role of PfSR1 in protecting P. falciparum from DNA damage and artemisinin exposure.

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The Role of UBR5 Mutations in the Pathogenesis of Mantle CELL Lymphoma

Blood ◽

10.1182/blood.v128.22.4124.4124 ◽

2016 ◽

Vol 128 (22) ◽

pp. 4124-4124

Author(s):

Olga Kutovaya ◽

Stacy Hung ◽

Hughes Christopher ◽

Randy D Gascoyne ◽

Morin Gregg ◽

...

Keyword(s):

Mass Spectrometry ◽

Cell Cycle ◽

Dna Damage ◽

Mantle Cell Lymphoma ◽

Dna Damage Response ◽

Cell Lymphoma ◽

Interacting Proteins ◽

Mantle Cell ◽

Damage Response

Abstract Intro: Mantle cell lymphoma (MCL) accounts for 6% of non-Hodgkin lymphomas and represents a particularly challenging disease with patient outcomes inferior to most other lymphoma subtypes. Using targeted capture sequencing of MCL biopsy samples, we recently reported frequent mutations (18%) in UBR5, a gene encoding an E3 ubiquitin-protein ligase that has not been previously implicated in lymphomagenesis. All mutations were clustered within 100bp in or around exon 58 of UBR5 and truncate the reading frame or change a key lysine residue. These mutations are predicted to result in the loss of the conserved cysteine residue in the HECT-domain, which is responsible for binding the ubiquitin co-factor. The recurrence and clustering of UBR5 mutations suggest their critical pathogenic involvement in a subgroup of MCL that might be therapeutically targetable. The aim of this study is to determine UBR5 mutation-associated proteome changes and altered cell signaling. Methods: As seen in MCL patients, mutations in exon 58 of UBR5 were introduced to three MCL cell lines (Granta-519, Jeko-1, and Mino) using the CRISPR-Cas9 genome engineering tool. First, mass spectrometry-based immunoprecipitation proteomics (IP-MS) was employed to identify differences in UBR5 interacting partners between UBR5 mutant and wildtype (WT) cells. Candidate UBR5 interacting proteins were validated by flow cytometry, western blotting, co-immunoprecipitation, and immunofluorescence. Next, global proteomes of UBR5 mutants and WT were analyzed by Tandem Mass Tag (TMT)-based mass spectrometry quantification to identify proteins with differential expression due to the UBR5 mutations. Results: The IP-MS analysis of WT vs UBR5 mutants revealed histone and cell cycle control proteins as candidate differential UBR5 interacting proteins (p<0.05). Particularly, histones H1, H4, and H2AFX, as well as the cell cycle genes CDC5L, BUB3, MAP4, RAD50 and CDK11B were identified as candidate UBR5 interacting partners. The global proteome analysis identified a set of differentially expressed genes (mutant vs wt; p<0.05) that are common among the MCL cell lines with the same direction of change. Gene ontology analysis of this set revealed DNA damage response, chromosome organization, and cell cycle response pathways as the predominant pathways affected. Moreover, our preliminary functional studies indicate constitutive phosphorylation of H2AFX in UBR5 mutants vs WT in line with the role of UBR5 in DNA damage response. Conclusions: Our results are consistent with UBR5 functioning as a key regulator of cell signalling and strongly suggest UBR5 as a novel regulator of histone modifications and DNA damage response. These findings provide an experimentally valid platform for further functional investigation and testing of target therapies for MCL harbouring UBR5 mutations. Disclosures No relevant conflicts of interest to declare.

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Characterization of the role of the DNA damage response pathway in parvoviral replication

10.32469/10355/15925 ◽

2012 ◽

Author(s):

Richard Adeyemi

Keyword(s):

Dna Damage ◽

Dna Damage Response ◽

Damage Response ◽

Dna Damage Response Pathway

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Faculty Opinions recommendation of A systems approach to delineate functions of paralogous transcription factors: role of the Yap family in the DNA damage response.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1103427.559478 ◽

2008 ◽

Author(s):

Jürg Bahler

Keyword(s):

Dna Damage ◽

Transcription Factors ◽

Dna Damage Response ◽

Systems Approach ◽

Damage Response

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DNA Damage Response in Multiple Myeloma: The Role of the Tumor Microenvironment

Cancers ◽

10.3390/cancers13030504 ◽

2021 ◽

Vol 13 (3) ◽

pp. 504

Author(s):

Takayuki Saitoh ◽

Tsukasa Oda

Keyword(s):

Multiple Myeloma ◽

Dna Damage ◽

Dna Repair ◽

Tumor Microenvironment ◽

Dna Damage Response ◽

Genetic Instability ◽

Dna Repair Mechanisms ◽

Damage Response ◽

Repair Mechanisms

Multiple myeloma (MM) is an incurable plasma cell malignancy characterized by genomic instability. MM cells present various forms of genetic instability, including chromosomal instability, microsatellite instability, and base-pair alterations, as well as changes in chromosome number. The tumor microenvironment and an abnormal DNA repair function affect genetic instability in this disease. In addition, states of the tumor microenvironment itself, such as inflammation and hypoxia, influence the DNA damage response, which includes DNA repair mechanisms, cell cycle checkpoints, and apoptotic pathways. Unrepaired DNA damage in tumor cells has been shown to exacerbate genomic instability and aberrant features that enable MM progression and drug resistance. This review provides an overview of the DNA repair pathways, with a special focus on their function in MM, and discusses the role of the tumor microenvironment in governing DNA repair mechanisms.

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Dissecting the Role of Thyrotropin in the DNA Damage Response in Human Thyrocytes after 131I, γ Radiation and H2O2

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/clinem/dgz185 ◽

2019 ◽

Vol 105 (3) ◽

pp. 839-853

Author(s):

Aglaia Kyrilli ◽

David Gacquer ◽

Vincent Detours ◽

Anne Lefort ◽

Frédéric Libert ◽

...

Keyword(s):

Gene Expression ◽

Cell Cycle ◽

Dna Damage ◽

Dna Damage Response ◽

Iodide Uptake ◽

Transcriptomic Response ◽

Uptake Inhibition ◽

Γ Radiation ◽

Damage Response

Abstract Background The early molecular events in human thyrocytes after 131I exposure have not yet been unravelled. Therefore, we investigated the role of TSH in the 131I-induced DNA damage response and gene expression in primary cultured human thyrocytes. Methods Following exposure of thyrocytes, in the presence or absence of TSH, to 131I (β radiation), γ radiation (3 Gy), and hydrogen peroxide (H2O2), we assessed DNA damage, proliferation, and cell-cycle status. We conducted RNA sequencing to profile gene expression after each type of exposure and evaluated the influence of TSH on each transcriptomic response. Results Overall, the thyrocyte responses following exposure to β or γ radiation and to H2O2 were similar. However, TSH increased 131I-induced DNA damage, an effect partially diminished after iodide uptake inhibition. Specifically, TSH increased the number of DNA double-strand breaks in nonexposed thyrocytes and thus predisposed them to greater damage following 131I exposure. This effect most likely occurred via Gα q cascade and a rise in intracellular reactive oxygen species (ROS) levels. β and γ radiation prolonged thyroid cell-cycle arrest to a similar extent without sign of apoptosis. The gene expression profiles of thyrocytes exposed to β/γ radiation or H2O2 were overlapping. Modulations in genes involved in inflammatory response, apoptosis, and proliferation were observed. TSH increased the number and intensity of modulation of differentially expressed genes after 131I exposure. Conclusions TSH specifically increased 131I-induced DNA damage probably via a rise in ROS levels and produced a more prominent transcriptomic response after exposure to 131I.

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The role of Schizosaccharomyces pombe Rad32, the Mre11 homologue, and other DNA damage response proteins in non-homologous end joining and telomere length maintenance

Nucleic Acids Research ◽

10.1093/nar/27.13.2655 ◽

1999 ◽

Vol 27 (13) ◽

pp. 2655-2661 ◽

Cited By ~ 50

Author(s):

S. Wilson ◽

N. Warr ◽

D. L. Taylor ◽

F. Z. Watts

Keyword(s):

Dna Damage ◽

Schizosaccharomyces Pombe ◽

Telomere Length ◽

Dna Damage Response ◽

End Joining ◽

Damage Response ◽

Non Homologous End Joining

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Role of H2AX in DNA damage response and human cancers

Mutation Research/Reviews in Mutation Research ◽

10.1016/j.mrrev.2008.08.003 ◽

2009 ◽

Vol 681 (2-3) ◽

pp. 180-188 ◽

Cited By ~ 53

Author(s):

Niloo Srivastava ◽

Sailesh Gochhait ◽

Peter de Boer ◽

Rameshwar N.K. Bamezai

Keyword(s):

Dna Damage ◽

Dna Damage Response ◽

Human Cancers ◽

Damage Response

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A novel role of the chromokinesin Kif4A in DNA damage response

Cell Cycle ◽

10.4161/cc.7.13.6130 ◽

2008 ◽

Vol 7 (13) ◽

pp. 2013-2020 ◽

Cited By ~ 37

Author(s):

Guikai Wu ◽

Longen Zhou ◽

Lily Khidr ◽

Xuning Emily Guo ◽

Wankee Kim ◽

...

Keyword(s):

Dna Damage ◽

Dna Damage Response ◽

Damage Response

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Nucleotide Excision Repair Protein Rad23 Regulates Cell Virulence Independent of Rad4 in Candida albicans

mSphere ◽

10.1128/msphere.00062-20 ◽

2020 ◽

Vol 5 (1) ◽

Cited By ~ 1

Author(s):

Jia Feng ◽

Shuangyan Yao ◽

Yansong Dong ◽

Jing Hu ◽

Malcolm Whiteway ◽

...

Keyword(s):

Dna Damage ◽

Candida Albicans ◽

Nucleotide Excision Repair ◽

Dna Damage Response ◽

Excision Repair ◽

Content Type ◽

Virulence Regulation ◽

Nucleotide Excision ◽

Damage Response

ABSTRACT In the pathogenic yeast Candida albicans, the DNA damage response contributes to pathogenicity by regulating cell morphology transitions and maintaining survival in response to DNA damage induced by reactive oxygen species (ROS) in host cells. However, the function of nucleotide excision repair (NER) in C. albicans has not been extensively investigated. To better understand the DNA damage response and its role in virulence, we studied the function of the Rad23 nucleotide excision repair protein in detail. The RAD23 deletion strain and overexpression strain both exhibit UV sensitivity, confirming the critical role of RAD23 in the nucleotide excision repair pathway. Genetic interaction assays revealed that the role of RAD23 in the UV response relies on RAD4 but is independent of RAD53, MMS22, and RAD18. RAD4 and RAD23 have similar roles in regulating cell morphogenesis and biofilm formation; however, only RAD23, but not RAD4, plays a negative role in virulence regulation in a mouse model. We found that the RAD23 deletion strain showed decreased survival in a Candida-macrophage interaction assay. Transcriptome sequencing (RNA-seq) and quantitative real-time PCR (qRT-PCR) data further revealed that RAD23, but not RAD4, regulates the transcription of a virulence factor, SUN41, suggesting a unique role of RAD23 in virulence regulation. Taking these observations together, our work reveals that the RAD23-related nucleotide excision pathway plays a critical role in the UV response but may not play a direct role in virulence. The virulence-related role of RAD23 may rely on the regulation of several virulence factors, which may give us further understanding about the linkage between DNA damage repair and virulence regulation in C. albicans. IMPORTANCE Candida albicans remains a significant threat to the lives of immunocompromised people. An understanding of the virulence and infection ability of C. albicans cells in the mammalian host may help with clinical treatment and drug discovery. The DNA damage response pathway is closely related to morphology regulation and virulence, as well as the ability to survive in host cells. In this study, we checked the role of the nucleotide excision repair (NER) pathway, the key repair system that functions to remove a large variety of DNA lesions such as those caused by UV light, but whose function has not been well studied in C. albicans. We found that Rad23, but not Rad4, plays a role in virulence that appears independent of the function of the NER pathway. Our research revealed that the NER pathway represented by Rad4/Rad23 may not play a direct role in virulence but that Rad23 may play a unique role in regulating the transcription of virulence genes that may contribute to the virulence of C. albicans.

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