RecBC酶将RecA蛋白不对称地加载到单链DNA上，且不依赖于χ序列，从而导致组成型重组激活。
The RecBC enzyme loads RecA protein onto ssDNA asymmetrically and independently of chi, resulting in constitutive recombination activation.
作者信息
Churchill J J, Anderson D G, Kowalczykowski S C
机构信息
Biochemistry and Molecular Biology Graduate Group, University of California, Davis, California 95616-8665 USA.
出版信息
Genes Dev. 1999 Apr 1;13(7):901-11. doi: 10.1101/gad.13.7.901.
Abstract
Double-strand DNA break repair and homologous recombination in Escherichia coli proceed by the RecBCD pathway, which is regulated by cis-acting elements known as chi sites. A crucial feature of this regulation is the RecBCD enzyme-directed loading of RecA protein specifically onto the 3'-terminal, chi-containing DNA strand. Here we show that RecBC enzyme (lacking the RecD subunit) loads RecA protein constitutively onto the 3'-terminal DNA strand, with no requirement for chi. This strand is preferentially utilized in homologous pairing reactions. We propose that RecA protein loading is a latent property of the RecBCD holoenzyme, which is normally blocked by the RecD subunit and is revealed following interaction with chi.
摘要
大肠杆菌中的双链DNA断裂修复和同源重组通过RecBCD途径进行，该途径受称为chi位点的顺式作用元件调控。这种调控的一个关键特征是RecBCD酶将RecA蛋白特异性加载到含chi的3'末端DNA链上。在这里，我们表明RecBC酶（缺乏RecD亚基）将RecA蛋白组成型地加载到3'末端DNA链上，而不需要chi。该链优先用于同源配对反应。我们提出，RecA蛋白加载是RecBCD全酶的一种潜在特性，通常被RecD亚基阻断，并在与chi相互作用后显现出来。
相似文献
1
The RecBC enzyme loads RecA protein onto ssDNA asymmetrically and independently of chi, resulting in constitutive recombination activation.RecBC酶将RecA蛋白不对称地加载到单链DNA上，且不依赖于χ序列，从而导致组成型重组激活。
Genes Dev. 1999 Apr 1;13(7):901-11. doi: 10.1101/gad.13.7.901.
2
Identification of the RecA protein-loading domain of RecBCD enzyme.RecBCD酶的RecA蛋白加载结构域的鉴定。
J Mol Biol. 2000 Mar 31;297(3):537-42. doi: 10.1006/jmbi.2000.3590.
3
The translocating RecBCD enzyme stimulates recombination by directing RecA protein onto ssDNA in a chi-regulated manner.易位的RecBCD酶通过以χ调控的方式将RecA蛋白引导到单链DNA上，从而刺激重组。
Cell. 1997 Jul 11;90(1):77-86. doi: 10.1016/s0092-8674(00)80315-3.
4
The RecD subunit of the Escherichia coli RecBCD enzyme inhibits RecA loading, homologous recombination, and DNA repair.大肠杆菌RecBCD酶的RecD亚基会抑制RecA装载、同源重组和DNA修复。
Proc Natl Acad Sci U S A. 2000 Jun 20;97(13):7399-404. doi: 10.1073/pnas.130192397.
5
Facilitated loading of RecA protein is essential to recombination by RecBCD enzyme.RecA蛋白的易化装载对于RecBCD酶介导的重组至关重要。
J Biol Chem. 2000 Apr 21;275(16):12261-5. doi: 10.1074/jbc.275.16.12261.
6
Role of the Escherichia coli recombination hotspot, chi, in RecABCD-dependent homologous pairing.大肠杆菌重组热点chi在RecABCD依赖的同源配对中的作用。
J Biol Chem. 1995 Jul 7;270(27):16360-70. doi: 10.1074/jbc.270.27.16360.
7
A RecA mutant, RecA(730), suppresses the recombination deficiency of the RecBC(1004)D-chi* interaction in vitro and in vivo.一种RecA突变体RecA(730)，在体外和体内均能抑制RecBC(1004)D-chi*相互作用的重组缺陷。
J Mol Biol. 2007 Feb 2;365(5):1314-25. doi: 10.1016/j.jmb.2006.10.090. Epub 2006 Nov 1.
8
In vitro reconstitution of homologous recombination reactions.同源重组反应的体外重建。
Experientia. 1994 Mar 15;50(3):204-15. doi: 10.1007/BF01924003.
9
A single mutation, RecB(D1080A,) eliminates RecA protein loading but not Chi recognition by RecBCD enzyme.单个突变RecB（D1080A）可消除RecA蛋白加载，但不影响RecBCD酶对Chi的识别。
J Biol Chem. 1999 Sep 17;274(38):27139-44. doi: 10.1074/jbc.274.38.27139.
10
Interaction with the recombination hot spot chi in vivo converts the RecBCD enzyme of Escherichia coli into a chi-independent recombinase by inactivation of the RecD subunit.在体内与重组热点chi相互作用，通过使RecD亚基失活，将大肠杆菌的RecBCD酶转化为不依赖chi的重组酶。
Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6249-53. doi: 10.1073/pnas.92.14.6249.
引用本文的文献
1
Trans-complementation by the RecB nuclease domain of RecBCD enzyme reveals new insight into RecA loading upon χ recognition.RecBCD 酶的 RecB 核酸酶结构域的反式互补揭示了 χ 识别后 RecA 加载的新见解。
Nucleic Acids Res. 2024 Mar 21;52(5):2578-2589. doi: 10.1093/nar/gkae007.
2
RecBCD enzyme: mechanistic insights from mutants of a complex helicase-nuclease.RecBCD 酶：从复杂解旋酶-核酸酶的突变体中获得的机制见解。
Microbiol Mol Biol Rev. 2023 Dec 20;87(4):e0004123. doi: 10.1128/mmbr.00041-23. Epub 2023 Dec 4.
3
Life, the genome and everything.生命、基因组与一切。
J Bacteriol. 2023 Dec 19;205(12):e0027223. doi: 10.1128/jb.00272-23. Epub 2023 Nov 29.
4
ppGpp and RNA-polymerase backtracking guide antibiotic-induced mutable gambler cells.ppGpp 和 RNA 聚合酶回溯引导抗生素诱导的可变性赌徒细胞。
Mol Cell. 2023 Apr 20;83(8):1298-1310.e4. doi: 10.1016/j.molcel.2023.03.003. Epub 2023 Mar 24.
5
CRISPR-Cas adaptation in Escherichia coli.CRISPR-Cas 系统在大肠杆菌中的适应性研究。
Biosci Rep. 2023 Mar 31;43(3). doi: 10.1042/BSR20221198.
6
Stress-Induced Mutagenesis, Gambler Cells, and Stealth Targeting Antibiotic-Induced Evolution.应激诱导突变、赌徒细胞和隐形靶向抗生素诱导进化。
mBio. 2022 Jun 28;13(3):e0107422. doi: 10.1128/mbio.01074-22. Epub 2022 Jun 6.
7
Insight into the biochemical mechanism of DNA helicases provided by bulk-phase and single-molecule assays.从体相和单分子检测中对 DNA 解旋酶生化机制的深入了解。
Methods. 2022 Aug;204:348-360. doi: 10.1016/j.ymeth.2021.12.002. Epub 2021 Dec 8.
8
Genomic mapping of DNA-repair reaction intermediates in living cells with engineered DNA structure-trap proteins.利用工程化 DNA 结构陷阱蛋白对活细胞中 DNA 修复反应中间体进行基因组定位。
Methods Enzymol. 2021;661:155-181. doi: 10.1016/bs.mie.2021.09.015. Epub 2021 Oct 29.
9
Efficient markerless integration of genes in the chromosome of probiotic E. coli Nissle 1917 by bacterial conjugation.通过细菌接合，高效地将基因无痕整合到益生菌 E. coli Nissle 1917 的染色体中。
Microb Biotechnol. 2022 May;15(5):1374-1391. doi: 10.1111/1751-7915.13967. Epub 2021 Nov 9.
10
Viral recombination systems limit CRISPR-Cas targeting through the generation of escape mutations.病毒重组系统通过产生逃逸突变限制了 CRISPR-Cas 的靶向。
Cell Host Microbe. 2021 Oct 13;29(10):1482-1495.e12. doi: 10.1016/j.chom.2021.09.001. Epub 2021 Sep 27.
本文引用的文献
1
ISOLATION AND CHARACTERIZATION OF RECOMBINATION-DEFICIENT MUTANTS OF ESCHERICHIA COLI K12.大肠杆菌K12重组缺陷突变体的分离与鉴定
Proc Natl Acad Sci U S A. 1965 Feb;53(2):451-9. doi: 10.1073/pnas.53.2.451.
2
A five-nucleotide sequence protects DNA from exonucleolytic degradation by AddAB, the RecBCD analogue of Bacillus subtilis.一个五核苷酸序列可保护DNA免受枯草芽孢杆菌的RecBCD类似物AddAB的核酸外切酶降解。
Mol Microbiol. 1998 Sep;29(6):1369-77. doi: 10.1046/j.1365-2958.1998.01018.x.
3
DNA strand exchange proteins: a biochemical and physical comparison.DNA链交换蛋白：生化与物理比较
Front Biosci. 1998 Jun 17;3:D570-603. doi: 10.2741/a304.
4
The 30-kDa C-terminal domain of the RecB protein is critical for the nuclease activity, but not the helicase activity, of the RecBCD enzyme from Escherichia coli.RecB蛋白的30 kDa C末端结构域对于来自大肠杆菌的RecBCD酶的核酸酶活性至关重要，但对于其解旋酶活性并非如此。
Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):981-6. doi: 10.1073/pnas.95.3.981.
5
Identification of the lactococcal exonuclease/recombinase and its modulation by the putative Chi sequence.乳酸乳球菌核酸外切酶/重组酶的鉴定及其假定的Chi序列对其的调控
Proc Natl Acad Sci U S A. 1998 Jan 20;95(2):626-31. doi: 10.1073/pnas.95.2.626.
6
Recombinational DNA repair: the RecF and RecR proteins limit the extension of RecA filaments beyond single-strand DNA gaps.重组DNA修复：RecF和RecR蛋白限制RecA丝在单链DNA缺口之外的延伸。
Cell. 1997 Oct 31;91(3):347-56. doi: 10.1016/s0092-8674(00)80418-3.
7
The translocating RecBCD enzyme stimulates recombination by directing RecA protein onto ssDNA in a chi-regulated manner.易位的RecBCD酶通过以χ调控的方式将RecA蛋白引导到单链DNA上，从而刺激重组。
Cell. 1997 Jul 11;90(1):77-86. doi: 10.1016/s0092-8674(00)80315-3.
8
The recombination hotspot Chi is recognized by the translocating RecBCD enzyme as the single strand of DNA containing the sequence 5'-GCTGGTGG-3'.重组热点Chi被易位的RecBCD酶识别为包含5'-GCTGGTGG-3'序列的单链DNA。
Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6706-11. doi: 10.1073/pnas.94.13.6706.
9
RecA protein: structure, function, and role in recombinational DNA repair.RecA蛋白：结构、功能及在重组DNA修复中的作用
Prog Nucleic Acid Res Mol Biol. 1997;56:129-223. doi: 10.1016/s0079-6603(08)61005-3.
10
Chi-activated RecBCD enzyme possesses 5'-->3' nucleolytic activity, but RecBC enzyme does not: evidence suggesting that the alteration induced by Chi is not simply ejection of the RecD subunit.被χ激活的RecBCD酶具有5'→3'核酸外切酶活性，但RecBC酶没有：证据表明χ诱导的改变不只是RecD亚基的排出。
Genes Cells. 1997 Feb;2(2):117-28. doi: 10.1046/j.1365-2443.1997.1130311.x.
Zotero 插件