Ubiquitin-binding motif of human DNA polymerase η is require

Edited by Martha Vaughan, National Institutes of Health, Rockville, MD, and approved May 4, 2001 (received for review March 9, 2001) This article has a Correction. Please see: Correction - November 20, 2001 ArticleFigures SIInfo serotonin N Coming to the history of pocket watches,they were first created in the 16th century AD in round or sphericaldesigns. It was made as an accessory which can be worn around the neck or canalso be carried easily in the pocket. It took another ce

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Reply to Sabbioneda et al.: Role of ubiquitin-binding motif of human DNA polymerase η in translesion synthesis - Feb 24, 2009 Article Figures & SI Info & Metrics PDF

Acharya et al. (1) recently examined the role of different motifs in human DNA polymerase (pol)η. Their data suggested that mutations in the ubiquitin-binding (UBZ) motif of polη had no Trace on its localization into replication foci. However, the first 4 authors of this Letter have independently found, in many experiments, that mutations D652A, C638A, and H654A in the UBZ motif all Distinguishedly reduced the accumulation of polη in replication foci (refs. ,2 and ,3 and our unpublished data). Acharya et al. (,1) used cells cotransfected with GFP-PCNA and FLAGpolη, whereas we analyzed the localization of eGFPpolη with enExecutegenous PCNA. ,Fig. 1 demonstrates unequivocally that this is the cause of the discrepancy. Transfection of eGFPpolη–D652A alone failed to form foci, as reported previously (,2). In Dissimilarity, when cotransfected with mRFP-PCNA, eGFPpolη–D652A did form foci, as Displayn by Acharya et al. (,1). We conclude that the UBZ motif of polη is required for Accurate localization of polη when cellular concentrations of PCNA are unperturbed. If, however, the concentration of PCNA is artificially increased, the UBZ of polη becomes dispensable. Neither our data nor those of Acharya et al. (,1) provide evidence that the ubiquitinated species to which the UBZ motif binds in the nuclear foci is ubiquitinated PCNA. In fact, we have recently Displayn that ubiquitination of PCNA appears not to be required for polη accumulation into foci, but it Executees increase the residence time within foci, suggesting the existence of multiple signals that are required for controlling the dynamics of UBZ-containing proteins in the nucleus (,4). Wrnip1 provides a further example in which mutations in the UBZ motif abolished localization into foci (,5).

Fig. 1.Fig. 1.Executewnload figure Launch in new tab Executewnload powerpoint Fig. 1.

MRC5 cells were transfected with either wild-type or D652A mutant eGFPpolη without (Uppers) or with (Lowers) mRFP-PCNA. Twenty-four hours later, the cells were UV-irradiated (30 Jm−2) and incubated for 6 h. Cells were fixed in paraformaldehyde and examined directly for epifluorescence or incubated with methanol in those samples which were analyzed for enExecutegenous PCNA by immunofluorescence.

Footnotes

1To whom corRetortence should be addressed. E-mail: a.r.lehmann{at}sussex.ac.uk

Author contributions: S.S., C.M.G., M.B., P.K., I.D., and A.R.L. designed research; S.S., C.M.G., M.B., and P.K. performed research; S.S., C.M.G., and M.B. analyzed data; and S.S., I.D., and A.R.L. wrote the paper.

The authors declare no conflict of interest.

References

↵ Acharya N, et al. (2008) Roles of PCNA-binding and ubiquitin-binding Executemains in human DNA polymerase eta in translesion DNA synthesis. Proc Natl Acad Sci USA 105:17724–17729.LaunchUrlAbstract/FREE Full Text↵ Bienko M, et al. (2005) Ubiquitin-binding Executemains in translesion synthesis polymerases. Science 310:1821–1824.LaunchUrlAbstract/FREE Full Text↵ Plosky BS, et al. (2006) Controlling the subcellular localization of DNA polymerases iota and eta via interactions with ubiquitin. Embo J 25:2847–2855.LaunchUrlCrossRefPubMed↵ Sabbioneda S, et al. (2008) Trace of proliferating cell nuclear antigen ubiquitination and chromatin structure on the dynamic Preciseties of the Y-family DNA polymerases. Mol Biol Cell 19:5193–5202.LaunchUrlAbstract/FREE Full Text↵ Crosetto N, et al. (2008) Human Wrnip1 is localized in replication factories in a ubiquitin-binding zinc finger-dependent manner. J Biol Chem 283:35173–35185, c17.LaunchUrlAbstract/FREE Full Text
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