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宿主介导的T4噬菌体DNA间断修复

Host-mediated repair of discontinuities in DNA from T4 bacteriophage.

作者信息

Carlson K, Lorkiewicz Z K, Kozinski A W

出版信息

J Virol. 1973 Aug;12(2):310-9. doi: 10.1128/JVI.12.2.310-319.1973.

Abstract

Discontinuities of T4 DNA which are caused by excision of UV-damaged areas, by decay of (32)P atoms, or which are present in DNA from rII(-)lig(am) (-) phage produced in a host nonpermissive for amber mutants are all repaired by bacterial enzymes after infection in the presence of chloramphenicol. Escherichia coli DNA polymerase I participates in the host-mediated repair, but an approximately 20-fold variation in the levels of host polynucleotide ligase does not affect either the kinetics or the extent of repair observed. Upon removal of chloramphenicol, host-repaired DNA from UV-irradiated phage undergoes a secondary cycle of breakage, which ultimately results in solubilization of most of the phage DNA. If the cells are co-infected with nonirradiated helper phage, the secondary breaks are repaired and the continuity of the polynucleotide chain is restored. The close coincidence in the extent of primary and secondary breakage suggests that phage-coded enzymes recognize and excise areas improperly repaired by the host. In contrast to host-mediated repair, repair mediated by rescuing phage probably restored functionality to the damaged DNA.

摘要

由紫外线损伤区域的切除、(32)P原子的衰变导致的T4 DNA的间断,或存在于在对琥珀突变体不允许的宿主中产生的rII(-)lig(am)(-)噬菌体的DNA中的间断,在氯霉素存在下感染后均由细菌酶修复。大肠杆菌DNA聚合酶I参与宿主介导的修复,但宿主多核苷酸连接酶水平约20倍的变化既不影响观察到的修复动力学也不影响修复程度。去除氯霉素后,来自紫外线照射噬菌体的宿主修复的DNA经历第二轮断裂,最终导致大部分噬菌体DNA溶解。如果细胞与未照射的辅助噬菌体共感染,第二轮断裂将被修复,多核苷酸链的连续性得以恢复。初次和二次断裂程度的紧密吻合表明噬菌体编码的酶识别并切除宿主修复不当的区域。与宿主介导的修复相反,拯救噬菌体介导的修复可能使受损DNA恢复功能。

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