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鉴定出鸭肠炎病毒编码与单纯疱疹病毒 1 的 UL15 蛋白同源的拼接基因。

Identification of a spliced gene from duck enteritis virus encoding a protein homologous to UL15 of herpes simplex virus 1.

机构信息

Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin 150001, China.

出版信息

Virol J. 2011 Apr 6;8:156. doi: 10.1186/1743-422X-8-156.

Abstract

BACKGROUND

In herpesviruses, UL15 homologue is a subunit of terminase complex responsible for cleavage and packaging of the viral genome into pre-assembled capsids. However, for duck enteritis virus (DEV), the causative agent of duck viral enteritis (DVE), the genomic sequence was not completely determined until most recently. There is limited information of this putative spliced gene and its encoding protein.

RESULTS

DEV UL15 consists of two exons with a 3.5 kilobases (kb) inron and transcribes into two transcripts: the full-length UL15 and an N-terminally truncated UL15.5. The 2.9 kb UL15 transcript encodes a protein of 739 amino acids with an approximate molecular mass of 82 kiloDaltons (kDa), whereas the UL15.5 transcript is 1.3 kb in length, containing a putative 888 base pairs (bp) ORF that encodes a 32 kDa product. We also demonstrated that UL15 gene belonged to the late kinetic class as its expression was sensitive to cycloheximide and phosphonoacetic acid. UL15 is highly conserved within the Herpesviridae, and contains Walker A and B motifs homologous to the catalytic subunit of the bacteriophage terminase as revealed by sequence analysis. Phylogenetic tree constructed with the amino acid sequences of 23 herpesvirus UL15 homologues suggests a close relationship of DEV to the Mardivirus genus within the Alphaherpesvirinae. Further, the UL15 and UL15.5 proteins can be detected in the infected cell lysate but not in the sucrose density gradient-purified virion when reacting with the antiserum against UL15. Within the CEF cells, the UL15 and/or UL15.5 localize(s) in the cytoplasm at 6 h post infection (h p. i.) and mainly in the nucleus at 12 h p. i. and at 24 h p. i., while accumulate(s) in the cytoplasm in the absence of any other viral protein.

CONCLUSIONS

DEV UL15 is a spliced gene that encodes two products encoded by 2.9 and 1.3 kb transcripts respectively. The UL15 is expressed late during infection. The coding sequences of DEV UL15 are very similar to those of alphaherpesviruses and most similar to the genus Mardivirus. The UL15 and/or UL15.5 accumulate(s) in the cytoplasm during early times post-infection and then are translocated to the nucleus at late times.

摘要

背景

在疱疹病毒中,UL15 同源物是末端酶复合物的一个亚基,负责将病毒基因组切割和包装到预先组装的衣壳中。然而,对于鸭肠炎病毒(DEV),即鸭病毒性肠炎(DVE)的病原体,直到最近才完全确定其基因组序列。关于这个假定的剪接基因及其编码蛋白的信息有限。

结果

DEV UL15 由两个外显子组成,内含子为 3.5 千碱基(kb),转录为两个转录本:全长 UL15 和 N 端截断的 UL15.5。2.9kb 的 UL15 转录本编码一个 739 个氨基酸的蛋白质,分子量约为 82 千道尔顿(kDa),而 UL15.5 转录本长 1.3kb,包含一个假定的 888 个碱基对(bp)ORF,编码一个 32kDa 的产物。我们还证明,UL15 基因属于晚期动力学类,因为其表达对环己酰亚胺和膦甲酸钠敏感。通过序列分析,UL15 在疱疹病毒科内高度保守,包含与噬菌体末端酶的催化亚基同源的 Walker A 和 B 基序。用 23 种疱疹病毒 UL15 同源物的氨基酸序列构建的系统发育树表明,DEV 与 Alphaherpesvirinae 中的 Mardivirus 属密切相关。此外,在感染细胞裂解物中可以检测到 UL15 和/或 UL15.5 蛋白,但在用针对 UL15 的抗血清反应时,不能在蔗糖密度梯度纯化的病毒粒子中检测到。在 CEF 细胞中,UL15 和/或 UL15.5 在感染后 6 小时(h p. i.)主要位于细胞质中,在 12 小时 p. i.和 24 小时 p. i.主要位于核内,而在没有任何其他病毒蛋白的情况下积累在细胞质中。

结论

DEV UL15 是一个剪接基因,分别由 2.9kb 和 1.3kb 转录本编码两种产物。UL15 在感染后期表达。DEV UL15 的编码序列与α疱疹病毒非常相似,与 Mardivirus 属最为相似。UL15 和/或 UL15.5 在感染后早期积累在细胞质中,然后在晚期转移到核内。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ac/3079670/4e0b05ff55c7/1743-422X-8-156-1.jpg

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