Ssb1 and Ssb2 cooperate to regulate mouse hematopoietic stem and progenitor cells by resolving replicative stress

Wei Shi, Therese Vu, Didier Boucher, Anna Biernacka, Jules Nde, Raj K. Pandita, Jasmin Straube, Glen M. Boyle, Fares Al-Ejeh, Purba Nag, Jessie Jeffery, Janelle L. Harris, Amanda L. Bain, Marta Grzelak, Magdalena Skrzypczak, Abhishek Mitra, Norbert Dojer, Nicola Crosetto, Nicole Cloonan, Olivier J. Becherel & 8 others John Finnie, Jeffrey R. Skaar, Carl R. Walkley, Tej K. Pandita, Malgorzata Rowicka-Kudlicka, Krzysztof Ginalski, Steven W. Lane, Kum Kum Khanna

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Hematopoietic stem and progenitor cells (HSPCs) are vulnerable to endogenous damage and defects in DNA repair can limit their function. The 2 single-stranded DNA (ssDNA) binding proteins SSB1 and SSB2 are crucial regulators of the DNA damage response; however, their overlapping roles during normal physiology are incompletely understood. We generated mice in which both Ssb1 and Ssb2 were constitutively or conditionally deleted. Constitutive Ssb1/Ssb2 double knockout (DKO) caused early embryonic lethality, whereas conditional Ssb1/Ssb2 double knockout (cDKO) in adult mice resulted in acute lethality due to bone marrow failure and intestinal atrophy featuring stem and progenitor cell depletion, a phenotype unexpected from the previously reported single knockout models of Ssb1 or Ssb2. Mechanistically, cDKO HSPCs showed altered replication fork dynamics, massive accumulation of DNA damage, genome-wide double-strand breaks enriched at Ssb-binding regions and CpG islands, together with the accumulation of R-loops and cytosolic ssDNA. Transcriptional profiling of cDKO HSPCs revealed the activation of p53 and interferon (IFN) pathways, which enforced cell cycling in quiescent HSPCs, resulting in their apoptotic death. The rapid cell death phenotype was reproducible in in vitro cultured cDKO-hematopoietic stem cells, which were significantly rescued by nucleotide supplementation or after depletion of p53. Collectively, Ssb1 and Ssb2 control crucial aspects of HSPC function, including proliferation and survival in vivo by resolving replicative stress to maintain genomic stability.

Original languageEnglish (US)
Pages (from-to)2471-2478
Number of pages8
JournalBlood
Volume129
Issue number18
DOIs
StatePublished - May 4 2017

Fingerprint

Hematopoietic Stem Cells
DNA
CpG Islands
Single-Stranded DNA
Physiology
DNA-Binding Proteins
Cell death
Stem cells
Interferons
Bone
Repair
Nucleotides
Genes
Chemical activation
Defects
DNA Damage
Stem Cells
Phenotype
Genomic Instability
DNA Repair

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Shi, W., Vu, T., Boucher, D., Biernacka, A., Nde, J., Pandita, R. K., ... Khanna, K. K. (2017). Ssb1 and Ssb2 cooperate to regulate mouse hematopoietic stem and progenitor cells by resolving replicative stress. Blood, 129(18), 2471-2478. https://doi.org/10.1182/blood-2016-06-725093

Ssb1 and Ssb2 cooperate to regulate mouse hematopoietic stem and progenitor cells by resolving replicative stress. / Shi, Wei; Vu, Therese; Boucher, Didier; Biernacka, Anna; Nde, Jules; Pandita, Raj K.; Straube, Jasmin; Boyle, Glen M.; Al-Ejeh, Fares; Nag, Purba; Jeffery, Jessie; Harris, Janelle L.; Bain, Amanda L.; Grzelak, Marta; Skrzypczak, Magdalena; Mitra, Abhishek; Dojer, Norbert; Crosetto, Nicola; Cloonan, Nicole; Becherel, Olivier J.; Finnie, John; Skaar, Jeffrey R.; Walkley, Carl R.; Pandita, Tej K.; Rowicka-Kudlicka, Malgorzata; Ginalski, Krzysztof; Lane, Steven W.; Khanna, Kum Kum.

In: Blood, Vol. 129, No. 18, 04.05.2017, p. 2471-2478.

Research output: Contribution to journalArticle

Shi, W, Vu, T, Boucher, D, Biernacka, A, Nde, J, Pandita, RK, Straube, J, Boyle, GM, Al-Ejeh, F, Nag, P, Jeffery, J, Harris, JL, Bain, AL, Grzelak, M, Skrzypczak, M, Mitra, A, Dojer, N, Crosetto, N, Cloonan, N, Becherel, OJ, Finnie, J, Skaar, JR, Walkley, CR, Pandita, TK, Rowicka-Kudlicka, M, Ginalski, K, Lane, SW & Khanna, KK 2017, 'Ssb1 and Ssb2 cooperate to regulate mouse hematopoietic stem and progenitor cells by resolving replicative stress', Blood, vol. 129, no. 18, pp. 2471-2478. https://doi.org/10.1182/blood-2016-06-725093
Shi, Wei ; Vu, Therese ; Boucher, Didier ; Biernacka, Anna ; Nde, Jules ; Pandita, Raj K. ; Straube, Jasmin ; Boyle, Glen M. ; Al-Ejeh, Fares ; Nag, Purba ; Jeffery, Jessie ; Harris, Janelle L. ; Bain, Amanda L. ; Grzelak, Marta ; Skrzypczak, Magdalena ; Mitra, Abhishek ; Dojer, Norbert ; Crosetto, Nicola ; Cloonan, Nicole ; Becherel, Olivier J. ; Finnie, John ; Skaar, Jeffrey R. ; Walkley, Carl R. ; Pandita, Tej K. ; Rowicka-Kudlicka, Malgorzata ; Ginalski, Krzysztof ; Lane, Steven W. ; Khanna, Kum Kum. / Ssb1 and Ssb2 cooperate to regulate mouse hematopoietic stem and progenitor cells by resolving replicative stress. In: Blood. 2017 ; Vol. 129, No. 18. pp. 2471-2478.
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abstract = "Hematopoietic stem and progenitor cells (HSPCs) are vulnerable to endogenous damage and defects in DNA repair can limit their function. The 2 single-stranded DNA (ssDNA) binding proteins SSB1 and SSB2 are crucial regulators of the DNA damage response; however, their overlapping roles during normal physiology are incompletely understood. We generated mice in which both Ssb1 and Ssb2 were constitutively or conditionally deleted. Constitutive Ssb1/Ssb2 double knockout (DKO) caused early embryonic lethality, whereas conditional Ssb1/Ssb2 double knockout (cDKO) in adult mice resulted in acute lethality due to bone marrow failure and intestinal atrophy featuring stem and progenitor cell depletion, a phenotype unexpected from the previously reported single knockout models of Ssb1 or Ssb2. Mechanistically, cDKO HSPCs showed altered replication fork dynamics, massive accumulation of DNA damage, genome-wide double-strand breaks enriched at Ssb-binding regions and CpG islands, together with the accumulation of R-loops and cytosolic ssDNA. Transcriptional profiling of cDKO HSPCs revealed the activation of p53 and interferon (IFN) pathways, which enforced cell cycling in quiescent HSPCs, resulting in their apoptotic death. The rapid cell death phenotype was reproducible in in vitro cultured cDKO-hematopoietic stem cells, which were significantly rescued by nucleotide supplementation or after depletion of p53. Collectively, Ssb1 and Ssb2 control crucial aspects of HSPC function, including proliferation and survival in vivo by resolving replicative stress to maintain genomic stability.",
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T1 - Ssb1 and Ssb2 cooperate to regulate mouse hematopoietic stem and progenitor cells by resolving replicative stress

AU - Shi, Wei

AU - Vu, Therese

AU - Boucher, Didier

AU - Biernacka, Anna

AU - Nde, Jules

AU - Pandita, Raj K.

AU - Straube, Jasmin

AU - Boyle, Glen M.

AU - Al-Ejeh, Fares

AU - Nag, Purba

AU - Jeffery, Jessie

AU - Harris, Janelle L.

AU - Bain, Amanda L.

AU - Grzelak, Marta

AU - Skrzypczak, Magdalena

AU - Mitra, Abhishek

AU - Dojer, Norbert

AU - Crosetto, Nicola

AU - Cloonan, Nicole

AU - Becherel, Olivier J.

AU - Finnie, John

AU - Skaar, Jeffrey R.

AU - Walkley, Carl R.

AU - Pandita, Tej K.

AU - Rowicka-Kudlicka, Malgorzata

AU - Ginalski, Krzysztof

AU - Lane, Steven W.

AU - Khanna, Kum Kum

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