Prokaryotic Argonautes(pAgos)provide bacteria and archaea with immunity against plasmids and viruses.Catalytically active pAgos utilize short oligonucleotides as guides to directly cleave foreign nucleic acids,while i...Prokaryotic Argonautes(pAgos)provide bacteria and archaea with immunity against plasmids and viruses.Catalytically active pAgos utilize short oligonucleotides as guides to directly cleave foreign nucleic acids,while inactive pAgos lacking catalytic residues employ auxiliary effectors,such as nonspecific nucleases,to trigger abortive infection upon detection of foreign nucleic acids.Here,we report a unique group of catalytically active pAgo proteins that frequently associate with a phos-pholipase D(PLD)family protein.We demonstrate that this particular system employs the catalytic center of the associated PLD protein rather than that of pAgo to restrict plasmid DNA,while interestingly,its immunity against a single-stranded DNA virus relies on the pAgo catalytic center and is enhanced by the PLD protein.We also find that this system selectively suppresses viral DNA propagation without inducing noticeable abortive infection outcomes.Moreover,the pAgo protein alone enhances gene editing,which is unexpectedly inhibited by the PLD protein.Our data highlight the ability of catalytically active pAgo proteins to employ auxiliary proteins to strengthen the targeted eradication of different genetic invaders and underline the trend of PLD nucleases to participate in host immunity.展开更多
Archaea represents the third domain of life, with the information-processing machineries more closely resembling those of eukaryotes than the machineries of the bacterial counterparts but sharing metabolic pathways wi...Archaea represents the third domain of life, with the information-processing machineries more closely resembling those of eukaryotes than the machineries of the bacterial counterparts but sharing metabolic pathways with organisms of Bacteria, the sister prokaryotic phylum. Archaeal organisms also possess unique features as revealed by genomics and genome comparisons and by biochemical characterization of prominent enzymes. Nevertheless, diverse genetic tools are required for in vivo experiments to verify these interesting discoveries. Considerable efforts have been devoted to the development of genetic tools for archaea ever since their discovery, and great progress has been made in the creation of archaeal genetic tools in the past decade. Versatile genetic toolboxes are now available for several archaeal models, among which Sulfolobus microorganisms are the only genus representing Crenarchaeota because all the remaining genera are from Euryarchaeota. Nevertheless, genetic tools developed for Sulfolobus are probably the most versatile among all archaeal models, and these include viral and plasmid shuttle vectors, conventional and novel genetic manipulation methods, CRISPR-based gene deletion and mutagenesis, and gene silencing, among which CRISPR tools have been reported only for Sulfolobus thus far. In this review, we summarize recent developments in all these useful genetic tools and discuss their possible application to research into archaeal biology by means of Sulfolobus models.展开更多
基金This work was supported by the Science&Technology Fundamental Resources Investigation Program(2022FY101100)the Strategic Priority Research Program of the Chinese Academy of Sciences(Precision seed design and breeding)(XDA24020101)+4 种基金the National Natural Science Foundation of China(Nos.31970544,31970545,32022003,32150020,32200057,and 32270092)the Youth Innovation Promotion Association of CAS(No.2020090)the China National Postdoctoral Program for Innovative Talents(BX20220331)the China Postdoctoral Science Foundation(2022M720160)the Special Research Assistant Program of Chinese Academy of Sciences(No.2023000056).
文摘Prokaryotic Argonautes(pAgos)provide bacteria and archaea with immunity against plasmids and viruses.Catalytically active pAgos utilize short oligonucleotides as guides to directly cleave foreign nucleic acids,while inactive pAgos lacking catalytic residues employ auxiliary effectors,such as nonspecific nucleases,to trigger abortive infection upon detection of foreign nucleic acids.Here,we report a unique group of catalytically active pAgo proteins that frequently associate with a phos-pholipase D(PLD)family protein.We demonstrate that this particular system employs the catalytic center of the associated PLD protein rather than that of pAgo to restrict plasmid DNA,while interestingly,its immunity against a single-stranded DNA virus relies on the pAgo catalytic center and is enhanced by the PLD protein.We also find that this system selectively suppresses viral DNA propagation without inducing noticeable abortive infection outcomes.Moreover,the pAgo protein alone enhances gene editing,which is unexpectedly inhibited by the PLD protein.Our data highlight the ability of catalytically active pAgo proteins to employ auxiliary proteins to strengthen the targeted eradication of different genetic invaders and underline the trend of PLD nucleases to participate in host immunity.
基金supported by the Danish Council of Independent Research (DFF-0602-02196, DFF-4181-00274, DFF-1323-00330)the Fundamental Research Funds for the Central Universities (2662015PX199)
文摘Archaea represents the third domain of life, with the information-processing machineries more closely resembling those of eukaryotes than the machineries of the bacterial counterparts but sharing metabolic pathways with organisms of Bacteria, the sister prokaryotic phylum. Archaeal organisms also possess unique features as revealed by genomics and genome comparisons and by biochemical characterization of prominent enzymes. Nevertheless, diverse genetic tools are required for in vivo experiments to verify these interesting discoveries. Considerable efforts have been devoted to the development of genetic tools for archaea ever since their discovery, and great progress has been made in the creation of archaeal genetic tools in the past decade. Versatile genetic toolboxes are now available for several archaeal models, among which Sulfolobus microorganisms are the only genus representing Crenarchaeota because all the remaining genera are from Euryarchaeota. Nevertheless, genetic tools developed for Sulfolobus are probably the most versatile among all archaeal models, and these include viral and plasmid shuttle vectors, conventional and novel genetic manipulation methods, CRISPR-based gene deletion and mutagenesis, and gene silencing, among which CRISPR tools have been reported only for Sulfolobus thus far. In this review, we summarize recent developments in all these useful genetic tools and discuss their possible application to research into archaeal biology by means of Sulfolobus models.
