Biochemical and cellular assays to assess the effects of acetylation on base excision repair enzymes

Shrabasti Roychoudhury, Suravi Pramanik, Hannah L. Harris, Kishor K Bhakat

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Protein posttranslational modifications (PTMs), including acetylation, have emerged as important regulators for controlling many cellular processes. DNA base excision repair (BER), a highly coordinated multistep cellular process, is primarily involved in the repair of both endogenous and drug-induced exogenous DNA base damages. BER relies on sequential recruitment and coordinated actions of multiple proteins. Increasing evidence suggests that acetylation of lysine residues of BER proteins facilitates fine-tuning of enzymatic activities, protein-protein interactions, and coordination of the steps in BER pathway. In this chapter, we describe detailed in vitro and in vivo approaches to examine the effect of acetylation on BER enzymes, focusing on the impact of acetylation of AP-endonuclease (APE1), a key enzyme in BER pathway, on its DNA damage repair activity, substrate-binding, and subcellular localization.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages191-206
Number of pages16
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1983
ISSN (Print)1064-3745

Fingerprint

Acetylation
DNA Repair
Enzymes
Proteins
DNA Damage
DNA-(Apurinic or Apyrimidinic Site) Lyase
Post Translational Protein Processing
Lysine
DNA
Pharmaceutical Preparations

Keywords

  • Acetylation
  • APE1
  • BER
  • Colony formation assay
  • Comet assay
  • DNA damage
  • DNA repair assay
  • EMSA
  • In vitro acetylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Roychoudhury, S., Pramanik, S., Harris, H. L., & Bhakat, K. K. (2019). Biochemical and cellular assays to assess the effects of acetylation on base excision repair enzymes. In Methods in Molecular Biology (pp. 191-206). (Methods in Molecular Biology; Vol. 1983). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9434-2_11

Biochemical and cellular assays to assess the effects of acetylation on base excision repair enzymes. / Roychoudhury, Shrabasti; Pramanik, Suravi; Harris, Hannah L.; Bhakat, Kishor K.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 191-206 (Methods in Molecular Biology; Vol. 1983).

Research output: Chapter in Book/Report/Conference proceedingChapter

Roychoudhury, S, Pramanik, S, Harris, HL & Bhakat, KK 2019, Biochemical and cellular assays to assess the effects of acetylation on base excision repair enzymes. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1983, Humana Press Inc., pp. 191-206. https://doi.org/10.1007/978-1-4939-9434-2_11
Roychoudhury S, Pramanik S, Harris HL, Bhakat KK. Biochemical and cellular assays to assess the effects of acetylation on base excision repair enzymes. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 191-206. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9434-2_11
Roychoudhury, Shrabasti ; Pramanik, Suravi ; Harris, Hannah L. ; Bhakat, Kishor K. / Biochemical and cellular assays to assess the effects of acetylation on base excision repair enzymes. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 191-206 (Methods in Molecular Biology).
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