Proteome-wide analysis of SUMO2 targets in response to pathological DNA replication stress in human cells

Sara Bursomanno; Petra Beli; Asif M Khan; Sheroy Minocherhomji; Sebastian Alexander Wagner; Simon Bekker-Jensen; Niels Mailand; Chuna Ram Choudhary; Ian D. Hickson; Ying Liu

doi:10.1016/j.dnarep.2014.10.011

Proteome-wide analysis of SUMO2 targets in response to pathological DNA replication stress in human cells

Sara Bursomanno, Petra Beli, Asif M Khan, Sheroy Minocherhomji, Sebastian Alexander Wagner, Simon Bekker-Jensen, Niels Mailand, Chuna Ram Choudhary, Ian D. Hickson, Ying Liu^*

^*Corresponding author for this work

28 Citations (Scopus)

Abstract

SUMOylation is a form of post-translational modification involving covalent attachment of SUMO (Small Ubiquitin-like Modifier) polypeptides to specific lysine residues in the target protein. In human cells, there are four SUMO proteins, SUMO1-4, with SUMO2 and SUMO3 forming a closely related subfamily. SUMO2/3, in contrast to SUMO1, are predominantly involved in the cellular response to certain stresses, including heat shock. Substantial evidence from studies in yeast has shown that SUMOylation plays an important role in the regulation of DNA replication and repair. Here, we report a proteomic analysis of proteins modified by SUMO2 in response to DNA replication stress in S phase in human cells. We have identified a panel of 22 SUMO2 targets with increased SUMOylation during DNA replication stress, many of which play key functions within the DNA replication machinery and/or in the cellular response to DNA damage. Interestingly, POLD3 was found modified most significantly in response to a low dose aphidicolin treatment protocol that promotes common fragile site (CFS) breakage. POLD3 is the human ortholog of POL32 in budding yeast, and has been shown to act during break-induced recombinational repair. We have also shown that deficiency of POLD3 leads to an increase in RPA-bound ssDNA when cells are under replication stress, suggesting that POLD3 plays a role in the cellular response to DNA replication stress. Considering that DNA replication stress is a source of genome instability, and that excessive replication stress is a hallmark of pre-neoplastic and tumor cells, our characterization of SUMO2 targets during a perturbed S-phase should provide a valuable resource for future functional studies in the fields of DNA metabolism and cancer biology.

Original language	English
Journal	DNA Repair
Volume	25
Pages (from-to)	84-96
Number of pages	13
ISSN	1568-7864
DOIs	https://doi.org/10.1016/j.dnarep.2014.10.011
Publication status	Published - Jan 2015

Keywords

Common fragile sites
DNA replication stress
Mass spectrometry
POLD3
SUMOylation

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.dnarep.2014.10.011

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title = "Proteome-wide analysis of SUMO2 targets in response to pathological DNA replication stress in human cells",

abstract = "SUMOylation is a form of post-translational modification involving covalent attachment of SUMO (Small Ubiquitin-like Modifier) polypeptides to specific lysine residues in the target protein. In human cells, there are four SUMO proteins, SUMO1-4, with SUMO2 and SUMO3 forming a closely related subfamily. SUMO2/3, in contrast to SUMO1, are predominantly involved in the cellular response to certain stresses, including heat shock. Substantial evidence from studies in yeast has shown that SUMOylation plays an important role in the regulation of DNA replication and repair. Here, we report a proteomic analysis of proteins modified by SUMO2 in response to DNA replication stress in S phase in human cells. We have identified a panel of 22 SUMO2 targets with increased SUMOylation during DNA replication stress, many of which play key functions within the DNA replication machinery and/or in the cellular response to DNA damage. Interestingly, POLD3 was found modified most significantly in response to a low dose aphidicolin treatment protocol that promotes common fragile site (CFS) breakage. POLD3 is the human ortholog of POL32 in budding yeast, and has been shown to act during break-induced recombinational repair. We have also shown that deficiency of POLD3 leads to an increase in RPA-bound ssDNA when cells are under replication stress, suggesting that POLD3 plays a role in the cellular response to DNA replication stress. Considering that DNA replication stress is a source of genome instability, and that excessive replication stress is a hallmark of pre-neoplastic and tumor cells, our characterization of SUMO2 targets during a perturbed S-phase should provide a valuable resource for future functional studies in the fields of DNA metabolism and cancer biology.",

keywords = "Common fragile sites, DNA replication stress, Mass spectrometry, POLD3, SUMOylation",

author = "Sara Bursomanno and Petra Beli and Khan, {Asif M} and Sheroy Minocherhomji and Wagner, {Sebastian Alexander} and Simon Bekker-Jensen and Niels Mailand and Choudhary, {Chuna Ram} and Hickson, {Ian D.} and Ying Liu",

year = "2015",

month = jan,

doi = "10.1016/j.dnarep.2014.10.011",

language = "English",

volume = "25",

pages = "84--96",

journal = "DNA Repair",

issn = "1568-7864",

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TY - JOUR

T1 - Proteome-wide analysis of SUMO2 targets in response to pathological DNA replication stress in human cells

AU - Bursomanno, Sara

AU - Beli, Petra

AU - Khan, Asif M

AU - Minocherhomji, Sheroy

AU - Wagner, Sebastian Alexander

AU - Bekker-Jensen, Simon

AU - Mailand, Niels

AU - Choudhary, Chuna Ram

AU - Hickson, Ian D.

AU - Liu, Ying

PY - 2015/1

Y1 - 2015/1

N2 - SUMOylation is a form of post-translational modification involving covalent attachment of SUMO (Small Ubiquitin-like Modifier) polypeptides to specific lysine residues in the target protein. In human cells, there are four SUMO proteins, SUMO1-4, with SUMO2 and SUMO3 forming a closely related subfamily. SUMO2/3, in contrast to SUMO1, are predominantly involved in the cellular response to certain stresses, including heat shock. Substantial evidence from studies in yeast has shown that SUMOylation plays an important role in the regulation of DNA replication and repair. Here, we report a proteomic analysis of proteins modified by SUMO2 in response to DNA replication stress in S phase in human cells. We have identified a panel of 22 SUMO2 targets with increased SUMOylation during DNA replication stress, many of which play key functions within the DNA replication machinery and/or in the cellular response to DNA damage. Interestingly, POLD3 was found modified most significantly in response to a low dose aphidicolin treatment protocol that promotes common fragile site (CFS) breakage. POLD3 is the human ortholog of POL32 in budding yeast, and has been shown to act during break-induced recombinational repair. We have also shown that deficiency of POLD3 leads to an increase in RPA-bound ssDNA when cells are under replication stress, suggesting that POLD3 plays a role in the cellular response to DNA replication stress. Considering that DNA replication stress is a source of genome instability, and that excessive replication stress is a hallmark of pre-neoplastic and tumor cells, our characterization of SUMO2 targets during a perturbed S-phase should provide a valuable resource for future functional studies in the fields of DNA metabolism and cancer biology.

AB - SUMOylation is a form of post-translational modification involving covalent attachment of SUMO (Small Ubiquitin-like Modifier) polypeptides to specific lysine residues in the target protein. In human cells, there are four SUMO proteins, SUMO1-4, with SUMO2 and SUMO3 forming a closely related subfamily. SUMO2/3, in contrast to SUMO1, are predominantly involved in the cellular response to certain stresses, including heat shock. Substantial evidence from studies in yeast has shown that SUMOylation plays an important role in the regulation of DNA replication and repair. Here, we report a proteomic analysis of proteins modified by SUMO2 in response to DNA replication stress in S phase in human cells. We have identified a panel of 22 SUMO2 targets with increased SUMOylation during DNA replication stress, many of which play key functions within the DNA replication machinery and/or in the cellular response to DNA damage. Interestingly, POLD3 was found modified most significantly in response to a low dose aphidicolin treatment protocol that promotes common fragile site (CFS) breakage. POLD3 is the human ortholog of POL32 in budding yeast, and has been shown to act during break-induced recombinational repair. We have also shown that deficiency of POLD3 leads to an increase in RPA-bound ssDNA when cells are under replication stress, suggesting that POLD3 plays a role in the cellular response to DNA replication stress. Considering that DNA replication stress is a source of genome instability, and that excessive replication stress is a hallmark of pre-neoplastic and tumor cells, our characterization of SUMO2 targets during a perturbed S-phase should provide a valuable resource for future functional studies in the fields of DNA metabolism and cancer biology.

KW - Common fragile sites

KW - DNA replication stress

KW - Mass spectrometry

KW - POLD3

KW - SUMOylation

U2 - 10.1016/j.dnarep.2014.10.011

DO - 10.1016/j.dnarep.2014.10.011

M3 - Journal article

C2 - 25497329

AN - SCOPUS:84917705873

SN - 1568-7864

VL - 25

SP - 84

EP - 96

JO - DNA Repair

JF - DNA Repair

ER -

Proteome-wide analysis of SUMO2 targets in response to pathological DNA replication stress in human cells

Abstract

Keywords

UN SDGs

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