title: The murine DNA glycosylase NEIL2 (mNEIL2) and human DNA polymerase beta bind microtubules in situ and in vitro.
abstract: 8-oxoguanine DNA glycosylase (OGG1), a major DNA repair enzyme in mammalian cells and a component of the base excision repair (BER) pathway, was recently shown to be associated with the microtubule network and the centriole at interphase and the spindle assembly at mitosis. In this study, we determined whether other participants in the BER pathway also bind microtubules in situ and in vitro. Purified recombinant human DNA polymerase beta (DNA Pol beta) and purified recombinant mNEIL2 were chemically conjugated to fluorochromes and photosensitive dyes and used in in situ localization and binding experiments. Results from in situ localization, microtubule co-precipitation and site-directed photochemical experiments showed that recombinant human DNA Pol beta and recombinant mNEIL2 associated with microtubules in situ and in vitro in a manner similar to that shown earlier for another BER pathway component, OGG1. Observations reported in this study suggest that these BER pathway components are microtubule-associated proteins (MAPs) themselves or utilize yet to be identified MAPs to bind microtubules in order to regulate their intracellular trafficking and activities during the cell cycle.
authors.affiliation: Department of Pharmacological Sciences, School of Medicine University Hospital and Medical Center, State University of New York Stony Brook, NY 11794 8651, USA.
authors.forename: Kimberly A
authors.initials: KA
authors.lastname: Conlon
authors.forename: Holly
authors.initials: H
authors.lastname: Miller
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Rosenquist
authors.forename: Dmitry O
authors.initials: DO
authors.lastname: Zharkov
authors.forename: Miguel
authors.initials: M
authors.lastname: Berrios
mesh_terms: D000818:Animals; D045647:DNA Glycosylases / Q000378:metabolism*; D019951:DNA Polymerase beta / Q000378:metabolism*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008870:Microtubules / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D019951:DNA Polymerase beta; D045647:DNA Glycosylases; C502613:Neil2 protein, mouse; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; C473642:NEIL2 protein, human