首页|The archaeal KEOPS complex possesses a functional Gon7 homolog and has an essential function independent of the cellular t6A modification level

The archaeal KEOPS complex possesses a functional Gon7 homolog and has an essential function independent of the cellular t6A modification level

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Kinase,putative Endopeptidase,and Other Proteins of Small size(KEOPS)is a multisubunit protein complex conserved in eukaryotes and archaea.It is composed of Pcc1,Kae1,Bud32,Cgi121,and Gon7 in eukaryotes and is primarily involved in N6-threonylcarbamoyl adenosine(t6A)modification of transfer RNAs(tRNAs).Recently,it was reported that KEOPS par-ticipates in homologous recombination(HR)repair in yeast.To characterize the KEOPS in archaea(aKEOPS),we conducted genetic and biochemical analyses of its encoding genes in the hyperthermophilic archaeon Saccharolobus islandicus.We show that aKEOPS also possesses five subunits,Pcc1,Kae1,Bud32,Cgi121,and Pcc1-like(or Gon7-like),just like eukaryotic KEOPS.Pcc1-like has physical interactions with Kae1 and Pcc1 and can mediate the monomerization of the dimeric sub-complex(Kae1-Pcc1-Pcc1-Kae1),suggesting that Pcc1-like is afunctional homolog of the eukaryotic Gon7 subunit.Strikingly,none of the genes encoding aKEOPS subunits,including Pcc1 and Pcc1-like,can be deleted in the wild type and in a t6A modification complementary strain named TsaKI,implying that the aKEOPS complex is essential for an additional cellular process in this archaeon.Knock-down of the Cgi121 subunit leads to severe growth retardance in the wild type that is partially rescued in TsaKI.These results suggest that aKEOPS plays an essential role independent of the cellular t6A modification level.In addition,archaeal Cgi121 possesses dsDNA-binding activity that relies on its tRNA 3'CCA tail binding module.Our study clarifies the subunit organization of archaeal KEOPS and suggests an origin of eukaryotic Gon7.The study also reveals a possible link between the function in t6A modification and the additional function,presumably HR.

archaeaDNA homologous recombination repairGon7KEOPSt6A modification

Pengju Wu、Qi Gan、Xuemei Zhang、Yunfeng Yang、Yuanxi Xiao、Qunxin She、Jinfeng Ni、Qihong Huang、Yulong Shen

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State Key Laboratory of Microbial Technology,CRISPR and Archaea Biology Research Center,Microbial Technology Institute,Shandong University,Qingdao,China

lab members of the CRISPR and Archaea Biology Research Centre for helpful discussionsNational Key Research and Development Program of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaState Key Laboratory of Microbial Technology

2020YFA090680031970546316700613190005531970119.31771380

2023

10.1002/mlf2.12051

微生物(英文)

微生物(英文)

ISSN：
年,卷(期)：2023.2(1)
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