RRC ID 80072
Author Koike M, Yamashita H, Yutoku Y, Koike A.
Title Molecular cloning, subcellular localization, and rapid recruitment to DNA damage sites of chicken Ku70.
Journal Sci Rep
Abstract Ku70 is a multifunctional protein with pivotal roles in DNA repair via non-homologous end-joining, V(D)J recombination, telomere maintenance, and neuronal apoptosis control. Nonetheless, its regulatory mechanisms remain elusive. Chicken Ku70 (GdKu70) cDNA has been previously cloned, and DT40 cells expressing it have significantly contributed to critical biological discoveries. GdKu70 features an additional 18 amino acids at its N-terminus compared to mammalian Ku70, the biological significance of which remains uncertain. Here, we show that the 5' flanking sequence of GdKu70 cDNA is not nearly encoded in the chicken genome. Notably, these 18 amino acids result from fusion events involving the NFE2L1 gene on chromosome 27 and the Ku70 gene on chromosome 1. Through experiments using newly cloned chicken Ku70 cDNA and specific antibodies, we demonstrated that Ku70 localizes within the cell nucleus as a heterodimer with Ku80 and promptly accumulates at DNA damage sites following injury. This suggests that the functions and spatiotemporal regulatory mechanisms of Ku70 in chickens closely resemble those in mammals. The insights and resources acquired will contribute to elucidate the various mechanisms by which Ku functions. Meanwhile, caution is advised when interpreting the previous numerous key studies that relied on GdKu70 cDNA and its expressing cells.
Volume 14(1)
Pages 1188
Published 2024-1-12
DOI 10.1038/s41598-024-51501-0
PII 10.1038/s41598-024-51501-0
PMID 38216643
PMC PMC10786929
MeSH Amino Acids / genetics Animals Antigens, Nuclear* / metabolism Chickens* / genetics Chickens* / metabolism Cloning, Molecular DNA Damage* / genetics DNA Repair DNA, Complementary DNA-Binding Proteins / metabolism Ku Autoantigen* / genetics Ku Autoantigen* / metabolism Mammals / metabolism
IF 3.998
Resource
Chicken / Quail M/O (PGC)