Carboxyl-terminal domain III of the delta' subunit of the DNA polymerase III holoenzyme binds delta - PubMed (original) (raw)
. 2001 Nov 2;276(44):40668-79.
doi: 10.1074/jbc.M106373200. Epub 2001 Aug 22.
Affiliations
- PMID: 11518714
- DOI: 10.1074/jbc.M106373200
Free article
Carboxyl-terminal domain III of the delta' subunit of the DNA polymerase III holoenzyme binds delta
M S Song et al. J Biol Chem. 2001.
Free article
Abstract
The delta and delta' subunits are essential components of the DNA polymerase III holoenzyme, required for assembly and function of the DnaX-complex clamp loader (tau2gammadeltadelta'chipsi). The x-ray crystal structure of delta' contains three structural domains (Guenther, B., Onrust, R., Sali, A., O'Donnell, M., and Kuriyan, J. (1997) Cell 91, 335-345). In this study, we localize the delta-binding domain of delta' to a carboxyl-terminal domain III by quantifying the interaction of delta with a series of delta' fusion proteins lacking specific domains. Purification and immobilization of the fusion proteins were facilitated by the inclusion of a tag containing hexahistidine and a short biotinylation sequence. Both NH2- and COOH-terminal-tagged full-length delta' were soluble and had specific activities comparable with that of native delta'. delta and delta' form a 1:1 heterodimer with a dissociation constant (K(D)) of 5 x 10(-7) m determined by equilibrium sedimentation. The K(D) determined by surface plasmon resonance was comparable. Domain III alone bound delta at an affinity comparable to that of wild type delta', whereas proteins lacking domain III did not bind delta. Using a panel of domain-specific anti-delta' monoclonal antibodies, we found that two of the domain III-specific monoclonal antibodies interfered with delta-delta' interaction and abolished the replication activity of DNA polymerase-III holoenzyme.
Similar articles
- Carboxyl-terminal domain III of the delta' subunit of DNA polymerase III holoenzyme binds DnaX and supports cooperative DnaX complex assembly.
Song MS, McHenry CS. Song MS, et al. J Biol Chem. 2001 Dec 28;276(52):48709-15. doi: 10.1074/jbc.M107936200. Epub 2001 Oct 17. J Biol Chem. 2001. PMID: 11606586 - The DnaX-binding subunits delta' and psi are bound to gamma and not tau in the DNA polymerase III holoenzyme.
Glover BP, McHenry CS. Glover BP, et al. J Biol Chem. 2000 Feb 4;275(5):3017-20. doi: 10.1074/jbc.275.5.3017. J Biol Chem. 2000. PMID: 10652279 - Clamp loader structure predicts the architecture of DNA polymerase III holoenzyme and RFC.
O'Donnell M, Jeruzalmi D, Kuriyan J. O'Donnell M, et al. Curr Biol. 2001 Nov 13;11(22):R935-46. doi: 10.1016/s0960-9822(01)00559-0. Curr Biol. 2001. PMID: 11719243 Review.
Cited by
- DNA Polymerase α Subunit Residues and Interactions Required for Efficient Initiation Complex Formation Identified by a Genetic Selection.
Lindow JC, Dohrmann PR, McHenry CS. Lindow JC, et al. J Biol Chem. 2015 Jul 3;290(27):16851-60. doi: 10.1074/jbc.M115.661090. Epub 2015 May 18. J Biol Chem. 2015. PMID: 25987558 Free PMC article. - Construction of bacteriophage phi29 DNA packaging motor and its applications in nanotechnology and therapy.
Lee TJ, Schwartz C, Guo P. Lee TJ, et al. Ann Biomed Eng. 2009 Oct;37(10):2064-81. doi: 10.1007/s10439-009-9723-0. Epub 2009 Jun 4. Ann Biomed Eng. 2009. PMID: 19495981 Free PMC article. Review. - Novel mechanism of hexamer ring assembly in protein/RNA interactions revealed by single molecule imaging.
Xiao F, Zhang H, Guo P. Xiao F, et al. Nucleic Acids Res. 2008 Nov;36(20):6620-32. doi: 10.1093/nar/gkn669. Epub 2008 Oct 21. Nucleic Acids Res. 2008. PMID: 18940870 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases