Points of recombination in Epstein-Barr virus (EBV) strain P3HR-1-derived heterogeneous DNA as indexes to EBV DNA recombinogenic events in vivo.
Points of recombination in Epstein-Barr virus (EBV) strain P3HR-1-derived heterogeneous DNA as indexes to EBV DNA recombinogenic events in vivo.
复制标题
EB 病毒 (EBV) 株 P3HR-1 衍生的异质 DNA 中的重组点作为体内 EBV DNA 重组事件的指标。
DOI：
10.1128/jvi.01036-08
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发表时间：
2008
影响因子：
5.4
通讯作者：
Sixbey,JohnW
中科院分区：
文献类型：
--
作者：
Ikuta,Kazufumi;Srinivas,ShamalaK;Schacker,Tim;Miyagi,Jun-ichi;Scott,RonaS;Sixbey,JohnW
EB病毒(EBV)P3HR-1株基因组的缺失和重排产生亚基因组感染性颗粒，与其他病毒系统中的缺陷干扰颗粒不同，在体外增强而不是限制EBV的复制。基于对EBV DNA片段BamHI W和BamHI Z异常并置的检测，在EBV相关人类疾病中有类似的异种(HET)DNA的报道，促使我们在核苷酸水平上确定由P3HR-1来源的HET DNA的四个组成片段形成的所有剩余重组关节。根据限制性内切酶图谱，我们选择了与BamHIM/B1和E/S融合片段相近和框架化的引物对，获得了BamHI M/B1和E/S融合片段。对聚合酶链式反应产物的测序显示，重组点在组成片段之间缺乏广泛的同源性区域。在感染人类免疫缺陷病毒的献血者的EBV阳性唾液样本中，用聚合酶链式反应检测到相同的重组连接，尽管在其他两个缺失的情况下，经常发现导致EBV重新激活的W/Z重排。淋巴细胞的体外感染同样表明，不是所有的三种het DNA重排都需要驻留在一个复合分子上。这些结果意味着重组过程中的精确度，该过程决定了基因组重排改变的基因片段的组成和调节。此外，体内EBV复制部位het DNA的表观频率与EBV重新激活的发病机制可能是一致的。
Deletions and rearrangements in the genome of Epstein-Barr virus (EBV) strain P3HR-1 generate subgenomic infectious particles that, unlike defective interfering particles in other viral systems, enhance rather than restrict EBV replication in vitro. Reports of comparable heterogeneous (het) DNA in EBV-linked human diseases, based on detection of an abnormal juxtaposition of EBV DNA fragments BamHI W and BamHI Z that disrupts viral latency, prompted us to determine at the nucleotide level all remaining recombination joints formed by the four constituent segments of P3HR-1-derived het DNA. Guided by endonuclease restriction maps, we chose PCR primer pairs that approximated and framed junctions creating the unique BamHI M/B1 and E/S fusion fragments. Sequencing of PCR products revealed points of recombination that lacked regions of extensive homology between constituent fragments. Identical recombination junctions were detected by PCR in EBV-positive salivary samples from human immunodeficiency virus-infected donors, although the W/Z rearrangement that induces EBV reactivation was frequently found in the absence of the other two. In vitro infection of lymphoid cells similarly indicated that not all three het DNA rearrangements need to reside on a composite molecule. These results connote a precision in the recombination process that dictates both composition and regulation of gene segments altered by genomic rearrangement. Moreover, the apparent frequency of het DNA at sites of EBV replication in vivo is consistent with a likely contribution to the pathogenesis of EBV reactivation.