Preparation of the gene targeted knockout mice for human premature aging diseases, Werner syndrome, and Rothmund-Thomson syndrome caused by the mutation of DNA helicases.

The RecQ helicases represent a subfamily of DNA helicases that are highly conserved in evolution. Loss of RecQ helicase function leads to a breakdown in the maintenance of genome integrity, in particular hyper-recombination. Germ-line defects in three of the five known human RecQ helicases give rise to defined genetic disorders associated with cancer predisposition and/or premature aging. These are Bloom's syndrome, Werner's syndrome and Rothmund–Thomson syndrome, which are caused by defects in the genes BLM, WRN and RECQ4 respectively. Here we review the properties of RecQ helicases in organisms from bacteria to humans, with an emphasis on the biochemical functions of these enzymes and the range of protein partners that they operate with. We will discuss models in which RecQ helicases are required to protect against replication fork demise, either through prevention of fork breakdown or restoration of productive DNA synthesis.

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Roles of the Bloom's syndrome helicase in the maintenance of genome stability

Biochemical Society Transactions ◽

10.1042/bst0331456 ◽

2005 ◽

Vol 33 (6) ◽

pp. 1456-1459 ◽

Cited By ~ 46

Author(s):

C.F. Cheok ◽

C.Z. Bachrati ◽

K.L. Chan ◽

C. Ralf ◽

L. Wu ◽

...

Keyword(s):

Genome Stability ◽

Protein A ◽

Sister Chromatid Exchanges ◽

Premature Aging ◽

Bloom's Syndrome ◽

Dna Helicases ◽

Bloom’S Syndrome ◽

Recombination Repair ◽

Rothmund Thomson Syndrome ◽

Chromatid Exchanges

The RecQ family of DNA helicases is highly conserved in evolution from bacteria to humans. Of the five known human RecQ family members, three (BLM, WRN and RECQ4, which cause Bloom's syndrome, Werner's syndrome and Rothmund–Thomson syndrome respectively) are mutated in distinct clinical disorders associated with cancer predisposition and/or premature aging. BLM forms part of a multienzyme complex including topoisomerase IIIα, replication protein A and a newly identified factor called BLAP75. Together, these proteins play a role in the resolution of DNA structures that arise during the process of homologous recombination repair. In the absence of BLM, cells show genomic instability and a high incidence of sister-chromatid exchanges. In addition to a DNA structure-specific helicase activity, BLM also catalyses Holliday-junction branch migration and the annealing of complementary single-stranded DNA molecules.

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The Role of Cellular Senescence in Werner Syndrome: Toward Therapeutic Intervention in Human Premature Aging

Annals of the New York Academy of Sciences ◽

10.1196/annals.1395.051 ◽

2007 ◽

Vol 1100 (1) ◽

pp. 455-469 ◽

Cited By ~ 48

Author(s):

T. DAVIS ◽

F. S. WYLLIE ◽

M. J. ROKICKI ◽

M. C. BAGLEY ◽

D. KIPLING

Keyword(s):

Cellular Senescence ◽

Therapeutic Intervention ◽

Werner Syndrome ◽

Premature Aging

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Human WRN is an intrinsic inhibitor of progerin, abnormal splicing product of lamin A

Scientific Reports ◽

10.1038/s41598-021-88325-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

So-mi Kang ◽

Min-Ho Yoon ◽

Su-Jin Lee ◽

Jinsook Ahn ◽

Sang Ah Yi ◽

...

Keyword(s):

Gene Expression ◽

Life Span ◽

Expression Analysis ◽

Gene Expression Analysis ◽

Werner Syndrome ◽

Genetic Disorder ◽

Dna Helicase ◽

Premature Aging ◽

Lamin A ◽

Short Life Span

AbstractWerner syndrome (WRN) is a rare progressive genetic disorder, caused by functional defects in WRN protein and RecQ4L DNA helicase. Acceleration of the aging process is initiated at puberty and the expected life span is approximately the late 50 s. However, a Wrn-deficient mouse model does not show premature aging phenotypes or a short life span, implying that aging processes differ greatly between humans and mice. Gene expression analysis of WRN cells reveals very similar results to gene expression analysis of Hutchinson Gilford progeria syndrome (HGPS) cells, suggesting that these human progeroid syndromes share a common pathological mechanism. Here we show that WRN cells also express progerin, an abnormal variant of the lamin A protein. In addition, we reveal that duplicated sequences of human WRN (hWRN) from exon 9 to exon 10, which differ from the sequence of mouse WRN (mWRN), are a natural inhibitor of progerin. Overexpression of hWRN reduced progerin expression and aging features in HGPS cells. Furthermore, the elimination of progerin by siRNA or a progerin-inhibitor (SLC-D011 also called progerinin) can ameliorate senescence phenotypes in WRN fibroblasts and cardiomyocytes, derived from WRN-iPSCs. These results suggest that progerin, which easily accumulates under WRN-deficient conditions, can lead to premature aging in WRN and that this effect can be prevented by SLC-D011.

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Telomerase Protects Werner Syndrome Lineage-Specific Stem Cells from Premature Aging

Stem Cell Reports ◽

10.1016/j.stemcr.2014.02.006 ◽

2014 ◽

Vol 2 (4) ◽

pp. 534-546 ◽

Cited By ~ 59

Author(s):

Hoi-Hung Cheung ◽

Xiaozhuo Liu ◽

Lucile Canterel-Thouennon ◽

Lu Li ◽

Catherine Edmonson ◽

...

Keyword(s):

Stem Cells ◽

Werner Syndrome ◽

Premature Aging

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WRN modulates translation by influencing mRNA export from the nucleus through its interaction with the export receptor NXF1 in HeLa cancer cell

10.21203/rs.3.rs-33612/v2 ◽

2020 ◽

Author(s):

Juan Manuel Iglesias-Pedraz ◽

Diego Matia Fossatti Jara ◽

Valeria Del Carmen Valle-Riestra Felice ◽

Sergio Rafael Cruz Visalaya ◽

Jose Antonio Ayala Felix ◽

...

Keyword(s):

Oxidative Stress ◽

Protein Synthesis ◽

Cancer Cells ◽

Nuclear Export ◽

Werner Syndrome ◽

Mrna Export ◽

Premature Aging ◽

Metabolic Shift ◽

Loss Of Function ◽

Global Protein Synthesis

Abstract Background The Werner syndrome protein (WRN) belongs to the RecQ family of helicases and its loss of function results in the premature aging disease Werner syndrome (WS). We previously demonstrated that an early cellular change induced by WRN depletion is a posttranscriptional decrease in the levels of enzymes involved in metabolic pathways that control macromolecular synthesis and protect from oxidative stress. This metabolic shift is tolerated by normal cells but causes mitochondria dysfunction and acute oxidative stress in rapidly growing cancer cells, thereby suppressing their proliferation.Results To identify the mechanism underlying this metabolic shift, we examined global protein synthesis and mRNA nucleocytoplasmic distribution after WRN knockdown. We determined that WRN depletion in HeLa cells attenuates global protein synthesis without affecting the level of key components of the mRNA export machinery. We further observed that WRN depletion affects the nuclear export of mRNAs and demonstrated that WRN directly interacts with mRNA and the mRNA export receptor Nuclear Export Factor 1 (NXF1).Conclusions Our findings suggest that WRN influences the export of mRNAs from the nucleus through its interaction with the NXF1 export receptor thereby affecting cellular proteostasis. In summary, we identified a new partner and a novel function of WRN, which is especially important for the proliferation of cancer cells.

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WRN modulates mRNA translation by influencing mRNA export from the nucleus through its interaction with the export receptor NXF1 in HeLa cancer cell

10.21203/rs.3.rs-33612/v1 ◽

2020 ◽

Author(s):

Juan Manuel Iglesias-Pedraz ◽

Diego Matia Fossatti Jara ◽

Valeria Del Carmen Valle-Riestra Felice ◽

Sergio Rafael Cruz Visalaya ◽

Jose Antonio Ayala Felix ◽

...

Keyword(s):

Oxidative Stress ◽

Protein Synthesis ◽

Cancer Cells ◽

Nuclear Export ◽

Werner Syndrome ◽

Mrna Export ◽

Mrna Translation ◽

Premature Aging ◽

Metabolic Shift ◽

Global Protein Synthesis

Abstract BackgroundThe Werner syndrome protein (WRN) belongs to the RecQ family of helicases and its loss of function results in the premature aging disease Werner syndrome (WS). We previously demonstrated that an early cellular change induced by WRN depletion is a posttranscriptional decrease in the levels of enzymes involved in metabolic pathways that control macromolecular synthesis and protect from oxidative stress. This metabolic shift is tolerated by normal cells but causes mitochondria dysfunction and acute oxidative stress in rapidly growing cancer cells, thereby suppressing their proliferation.Results To identify the mechanism underlying this metabolic shift, we examined global protein synthesis and mRNA nucleocytoplasmic distribution after WRN knockdown. We determined that WRN depletion in HeLa cells attenuates global protein synthesis without affecting the level of key components of the mRNA export machinery. We further observed that WRN depletion affects the nuclear export of mRNAs and demonstrated that WRN directly interacts with mRNA and the mRNA export receptor Nuclear Export Factor 1 (NXF1).Conclusions Our findings suggest that WRN influences the export of mRNAs from the nucleus through its interaction with the NXF1 export receptor thereby affecting cellular proteostasis. In summary, we identified a new partner and a novel function of WRN, which is especially important for the proliferation of cancer cells.

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WRN modulates translation by influencing nuclear mRNA export in HeLa cancer cells

10.21203/rs.3.rs-33612/v5 ◽

2020 ◽

Author(s):

Juan Manuel Iglesias-Pedraz ◽

Diego Matia Fossatti Jara ◽

Valeria Del Carmen Valle-Riestra Felice ◽

Sergio Rafael Cruz Visalaya ◽

Jose Antonio Ayala Felix ◽

...

Keyword(s):

Oxidative Stress ◽

Protein Synthesis ◽

Cancer Cells ◽

Nuclear Export ◽

Werner Syndrome ◽

Mrna Export ◽

Premature Aging ◽

Metabolic Shift ◽

Loss Of Function ◽

Global Protein Synthesis

Abstract BackgroundThe Werner syndrome protein (WRN) belongs to the RecQ family of helicases and its loss of function results in the premature aging disease Werner syndrome (WS). We previously demonstrated that an early cellular change induced by WRN depletion is a posttranscriptional decrease in the levels of enzymes involved in metabolic pathways that control macromolecular synthesis and protect from oxidative stress. This metabolic shift is tolerated by normal cells but causes mitochondria dysfunction and acute oxidative stress in rapidly growing cancer cells, thereby suppressing their proliferation.ResultsTo identify the mechanism underlying this metabolic shift, we examined global protein synthesis and mRNA nucleocytoplasmic distribution after WRN knockdown. We determined that WRN depletion in HeLa cells attenuates global protein synthesis without affecting the level of key components of the mRNA export machinery. We further observed that WRN depletion affects the nuclear export of mRNAs and demonstrated that WRN interacts with mRNA and the Nuclear RNA Export Factor 1 (NXF1).ConclusionsOur findings suggest that WRN influences the export of mRNAs from the nucleus through its interaction with the NXF1 export receptor thereby affecting cellular proteostasis. In summary, we identified a new partner and a novel function of WRN, which is especially important for the proliferation of cancer cells.

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A genome-wide CRISPR-based screen identifies KAT7 as a driver of cellular senescence

Science Translational Medicine ◽

10.1126/scitranslmed.abd2655 ◽

2021 ◽

Vol 13 (575) ◽

pp. eabd2655

Author(s):

Wei Wang ◽

Yuxuan Zheng ◽

Shuhui Sun ◽

Wei Li ◽

Moshi Song ◽

...

Keyword(s):

Cellular Senescence ◽

Werner Syndrome ◽

Embryonic Stem ◽

Accelerated Aging ◽

Premature Aging ◽

Mesenchymal Precursor Cells ◽

Genome Wide ◽

A Genome ◽

Progeria Syndrome ◽

Hutchinson Gilford Progeria Syndrome

Understanding the genetic and epigenetic bases of cellular senescence is instrumental in developing interventions to slow aging. We performed genome-wide CRISPR-Cas9–based screens using two types of human mesenchymal precursor cells (hMPCs) exhibiting accelerated senescence. The hMPCs were derived from human embryonic stem cells carrying the pathogenic mutations that cause the accelerated aging diseases Werner syndrome and Hutchinson-Gilford progeria syndrome. Genes whose deficiency alleviated cellular senescence were identified, including KAT7, a histone acetyltransferase, which ranked as a top hit in both progeroid hMPC models. Inactivation of KAT7 decreased histone H3 lysine 14 acetylation, repressed p15INK4b transcription, and alleviated hMPC senescence. Moreover, lentiviral vectors encoding Cas9/sg-Kat7, given intravenously, alleviated hepatocyte senescence and liver aging and extended life span in physiologically aged mice as well as progeroid Zmpste24−/− mice that exhibit a premature aging phenotype. CRISPR-Cas9–based genetic screening is a robust method for systematically uncovering senescence genes such as KAT7, which may represent a therapeutic target for developing aging interventions.

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Human Fbh1 helicase contributes to genome maintenance via pro- and anti-recombinase activities

The Journal of Cell Biology ◽

10.1083/jcb.200812138 ◽

2009 ◽

Vol 186 (5) ◽

pp. 655-663 ◽

Cited By ~ 66

Author(s):

Kasper Fugger ◽

Martin Mistrik ◽

Jannie Rendtlew Danielsen ◽

Christoffel Dinant ◽

Jacob Falck ◽

...

Keyword(s):

Small Interfering Rna ◽

Ectopic Expression ◽

Dna Lesions ◽

Premature Aging ◽

Genome Integrity ◽

Helicase Activity ◽

Genome Maintenance ◽

Dna Helicases ◽

Replication Block ◽

Interfering Rna

Homologous recombination (HR) is essential for faithful repair of DNA lesions yet must be kept in check, as unrestrained HR may compromise genome integrity and lead to premature aging or cancer. To limit unscheduled HR, cells possess DNA helicases capable of preventing excessive recombination. In this study, we show that the human Fbh1 (hFbh1) helicase accumulates at sites of DNA damage or replication stress in a manner dependent fully on its helicase activity and partially on its conserved F box. hFbh1 interacted with single-stranded DNA (ssDNA), the formation of which was required for hFbh1 recruitment to DNA lesions. Conversely, depletion of endogenous Fbh1 or ectopic expression of helicase-deficient hFbh1 attenuated ssDNA production after replication block. Although elevated levels of hFbh1 impaired Rad51 recruitment to ssDNA and suppressed HR, its small interfering RNA–mediated depletion increased the levels of chromatin-associated Rad51 and caused unscheduled sister chromatid exchange. Thus, by possessing both pro- and anti-recombinogenic potential, hFbh1 may cooperate with other DNA helicases in tightly controlling cellular HR activity.

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