Molecular cloning, subcellular localization, and rapid recruitment to DNA damage sites of chicken Ku70.

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

Original Publication: London : Nature Publishing Group, copyright 2011-

Antigens, Nuclear*/metabolism ; Chickens*/genetics ; Chickens*/metabolism ; DNA Damage*/genetics ; Ku Autoantigen*/genetics ; Ku Autoantigen*/metabolism; Animals ; Amino Acids/genetics ; Cloning, Molecular ; DNA Repair ; DNA, Complementary ; DNA-Binding Proteins/metabolism ; Mammals/metabolism

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.
(© 2024. The Author(s).)

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JP22K06036 Japan Society for the Promotion of Science

0 (Amino Acids)
0 (Antigens, Nuclear)
0 (DNA, Complementary)
0 (DNA-Binding Proteins)
EC 4.2.99.- (Ku Autoantigen)

Date Created: 20240112 Date Completed: 20240206 Latest Revision: 20240206

20240206

PMC10786929

10.1038/s41598-024-51501-0

38216643