CENPA is regulated by both DDR and SAC components and is required for recruitment of RAD-51 to the nuclear periphery and efficient DNA damage repair.

<p>(A) Proliferative zones of wild-type, <i>mad-1(gk2)</i> and <i>atr(tm853)</i> worms treated with HU and stained for CENPA (red), Mab414 (NPC) (green) and DAPI (blue). (B) Quantification of the average fluorescence intensity of CENPA, MAb414, and DAPI across the nucleus length binned in 10% increments-HU for wild type, <i>mad-1(gk2)</i>and <i>atr(tm853)</i>, and +HU for the above genotypes (n≥30). (C) Quantification of the average fluorescence intensity of CENPA and Mab414 across the nucleus length binned in 10% increments for <i>mad-2</i>(RNAi), <i>mad-3(ok1580)</i>, <i>bub-</i>3(RNAi), <i>knl-1</i>(RNAi) and <i>fzy-1(av15)</i> in the presence of HU (n≥30). (D) Percent of nuclei that contain at least 1 RAD-51 focus—HU or 0, 2, 4, 6, or 16 hours after release from 5mM HU with <i>cenpa</i>(RNAi); n≥10. Scale bars 10μm. (E) SIM images of a single nucleus from wild-type, <i>mad-1(gk2)</i> and <i>cenpa</i>(RNAi) worms treated with HU. Next to each image is the average number of RAD-51 foci observed in each genotype, as well as the average area of each RAD-51 focus (n = 4 germ lines). Images represent a projection of 3 z sections. Graph indicates the distance in nanometers between NPC and RAD-51 foci in wild-type, <i>mad-1(gk2)</i> and <i>cenpa</i>(RNAi) worms after HU. Scale bar 2 μm. Error bars indicate SEM. ***p<0.0001