CRISPR-Associated Primase-Polymerases are implicated in prokaryotic CRISPR-Cas adaptation.
CRISPR-Associated Primase-Polymerases are implicated in prokaryotic CRISPR-Cas adaptation.
复制标题
DOI：
10.1038/s41467-021-23535-9
复制
发表时间：
2021-06-17
影响因子：
16.6
通讯作者：
Doherty AJ
中科院分区：
文献类型：
--
作者：
Zabrady K;Zabrady M;Kolesar P;Li AWH;Doherty AJ
CRISPR-Cas途径为原核生物提供了针对外源遗传元件（包括DNA和质粒）的获得性“免疫力”。尽管许多与CRISPR-Cas机制相关的蛋白质已被表征，但一些必需的酶仍然难以捉摸。遗传学研究已经在一些CRISPR-Cas系统中涉及宿主DNA聚合酶，但尚未发现CRISPR特异性复制酶。我们已经鉴定并表征了一系列原核生物中的CRISPR相关引物聚合酶（CAPP）家族，这些原核生物与Cas 1和Cas 2操纵相关。CAPP属于引物聚合酶（Prim-Pol）复制酶超家族，其在维持基因组稳定性的各种DNA修复和复制途径中操作。在这里，我们检测了来自IIIA型和IIIB型CRISPR-Cas系统的细菌CAPP同源物的DNA合成活性，并确定它们具有一系列复制酶活性，包括DNA引发、聚合和链置换。我们证明，CAPPs的operonically相关的合作伙伴，Cas 1和Cas 2，形成一个复杂的，具有间隔区整合活性。我们发现CAPP与Cas蛋白物理结合，形成定制的CRISPR-Cas复合物。最后，我们提出了CAPP活动如何与其合作伙伴一起在CRISPR-Cas适应中发挥关键作用。CAPP是与CRISPR-Cas操纵子相关的推定引物聚合酶。在这里，作者显示了CAPPs与Cas 1和Cas 2的遗传和物理关联，它们作为DNA依赖性DNA引发酶和DNA聚合酶的能力，以及与CAPP相邻的Cas 1-Cas 2复合物具有真正的间隔区整合活性。
CRISPR-Cas pathways provide prokaryotes with acquired “immunity” against foreign genetic elements, including phages and plasmids. Although many of the proteins associated with CRISPR-Cas mechanisms are characterized, some requisite enzymes remain elusive. Genetic studies have implicated host DNA polymerases in some CRISPR-Cas systems but CRISPR-specific replicases have not yet been discovered. We have identified and characterised a family of CRISPR-Associated Primase-Polymerases (CAPPs) in a range of prokaryotes that are operonically associated with Cas1 and Cas2. CAPPs belong to the Primase-Polymerase (Prim-Pol) superfamily of replicases that operate in various DNA repair and replication pathways that maintain genome stability. Here, we characterise the DNA synthesis activities of bacterial CAPP homologues from Type IIIA and IIIB CRISPR-Cas systems and establish that they possess a range of replicase activities including DNA priming, polymerisation and strand-displacement. We demonstrate that CAPPs operonically-associated partners, Cas1 and Cas2, form a complex that possesses spacer integration activity. We show that CAPPs physically associate with the Cas proteins to form bespoke CRISPR-Cas complexes. Finally, we propose how CAPPs activities, in conjunction with their partners, may function to undertake key roles in CRISPR-Cas adaptation. CAPPs are putative Primase-Polymerases associated with CRISPR-Cas operons. Here, the authors show CAPPs genetic and physical association with Cas1 and Cas2, their capacity to function as DNA-dependent DNA primases and DNA polymerases, and that Cas1-Cas2 complex adjacent to CAPP has bona fide spacer integration activity.
登录
查看更多内容
影响因子：
14.9
作者：
Guilliam TA;Keen BA;Brissett NC;Doherty AJ
通讯作者：
Doherty AJ
影响因子：
14.9
作者：
Garrett, Sandra;Shiimori, Masami;Terns, Michael P.
通讯作者：
Terns, Michael P.
影响因子：
16.8
作者：
Gong, CL;Bongiorno, P;Glickman, MS
通讯作者：
Glickman, MS
影响因子：
56.9
作者：
Della, M;Palmbos, PL;Doherty, AJ
通讯作者：
Doherty, AJ
影响因子：
3.6
作者：
Babu M;Beloglazova N;Flick R;Graham C;Skarina T;Nocek B;Gagarinova A;Pogoutse O;Brown G;Binkowski A;Phanse S;Joachimiak A;Koonin EV;Savchenko A;Emili A;Greenblatt J;Edwards AM;Yakunin AF
通讯作者：
Yakunin AF