REV1 protein interacts with PCNA: significance of the REV1 BRCT domain in vitro and in vivo

REV1 protein, a eukaryotic member of the Y family of DNA polymerases, is involved in the tolerance of DNA damage by translesion DNA synthesis. It is unclear how REV1 is recruited to replication foci in cells. Here, we report that mouse REV1 can bind directly to PCNA and that monoubiquitylation of PCNA enhances this interaction. The interaction between REV1 protein and PCNA requires a functional BRCT domain located near the N terminus of the former protein. Deletion or mutational inactivation of the BRCT domain abolishes the targeting of REV1 to replication foci in unirradiated cells, but not in UV-irradiated cells. In vivo studies in both chicken DT40 cells and yeast directly support the requirement of the BRCT domain of REV1 for cell survival and DNA damage-induced mutagenesis.     

The 2nd US–Japan DNA Repair Meeting, Honolulu, Hawaii, June 4–8, 2004

The 2nd US–Japan DNA Repair Meeting convened at the JW Marriott Ihilani Hotel, outside Honolulu, Hawaii, from June 4–8, 2004. In keeping with the tradition of US–Japan conferences the meeting was modest in size comprising 25 participants from each country. The program featured platform presentations from each participant, with lots of time devoted to discussion of groups of related talks. A novel feature of the meeting was the absence of formally designated and previously announced titles for sessions and talks, providing a level of informality that promoted relaxed interactions. Discussion was gratifyingly brisk and informative throughout and was considered to be a highlight of the meeting. All sessions were chaired by the program planners Errol Friedberg and Sam Wilson, who did not present formal talks. The following pages comprise summations of the talks presented, organized into primary topic themes.     

Singlet CH domain containing human multidomain proteins: an inventory

The actin cytoskeleton presents the basic force in processes such as cytokinesis, endocytosis, vesicular trafficking and cell migration. Here, we list 30 human singlet CH (calpononin homology/actin binding) containing multidomain molecules, each encoded by one gene. We show the domain distributions as given by the SMART program. These mosaic proteins organize geographically the placement of selected proteins in proximity within the cell. In most instances, their precise location, their actin binding capacity by way of the singlet CH (or by other domains?) and their physiological functions need further elucidation. A dendrogram based solely on the relationship for the human singlet CH domains (in terms of AA sequences) for the various molecules that possess the domain, implies that the singlet descended from a common ancestor which in turn sprouted three main branches of protein products. Each branch bifurcated multiple times thus accounting for a cornucopia of products. Wherever, additional (unassigned), highly homologous regions exist in related proteins (e.g., in LIM and LMO7 or in Tangerin and EH/BP1), these unrecognized domain regions await assignment as specific functional domains. Frequently genes coding multidomain proteins duplicated. The varying modular nature within multidomain proteins should have accelerated evolutionary changes to a degree not feasible to achieve by means of mere post-duplication mutational changes.