title: Catalytic activities of Werner protein are affected by adduction with 4-hydroxy-2-nonenal.
abstract: 4-Hydroxy-2-nonenal (HNE) is a reactive α,β-unsaturated aldehyde generated during oxidative stress and subsequent peroxidation of polyunsaturated fatty acids. Here, Werner protein (WRN) was identified as a novel target for modification by HNE. Werner syndrome arises through mutations in the WRN gene that encodes the RecQ DNA helicase which is critical for maintaining genomic stability. This hereditary disease is associated with chromosomal instability, premature aging and cancer predisposition. WRN appears to participate in the cellular response to oxidative stress and cells devoid of WRN display elevated levels of oxidative DNA damage. We demonstrated that helicase/ATPase and exonuclease activities of HNE-modified WRN protein were inhibited both in vitro and in immunocomplexes purified from the cell extracts. Sites of HNE adduction in human WRN were identified at Lys577, Cys727, His1290, Cys1367, Lys1371 and Lys1389. We applied in silico modeling of the helicase and RQC domains of WRN protein with HNE adducted to Lys577 and Cys727 and provided a potential mechanism of the observed deregulation of the protein catalytic activities. In light of the obtained results, we postulate that HNE adduction to WRN is a post-translational modification, which may affect WRN conformational stability and function, contributing to features and diseases associated with premature senescence.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
authors.forename: Jolanta
authors.initials: J
authors.lastname: Czerwińska
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
authors.forename: Jarosław
authors.initials: J
authors.lastname: Poznański
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
authors.forename: Janusz
authors.initials: J
authors.lastname: Dębski
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
authors.forename: Zuzanna
authors.initials: Z
authors.lastname: Bukowy
authors.affiliation: National Institute on Aging, National Institutes of Health, 251 Bayview Blvd, Baltimore, MD 21224, USA.
authors.forename: Vilhelm A
authors.initials: VA
authors.lastname: Bohr
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland Institute of Genetics and Biotechnology, University of Warsaw, Pawińskiego 5a, 02-106 Warsaw, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Tudek
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland elasp@ibb.waw.pl.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Speina
mesh_terms: D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000447:Aldehydes / Q000032:analysis / Q000737:chemistry*; D000818:Animals; D055162:Biocatalysis; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D005090:Exodeoxyribonucleases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D053484:RecQ Helicases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D000071657:Werner Syndrome Helicase
chemical_list: D000447:Aldehydes; D004247:DNA; D005090:Exodeoxyribonucleases; D000251:Adenosine Triphosphatases; D053484:RecQ Helicases; C098785:WRN protein, human; D000071657:Werner Syndrome Helicase; C027576:4-hydroxy-2-nonenal