Martinez-Guzman DeeAnn, Rickabaugh Tammy, Wu Ting-Ting, Brown Helen, Cole Steven, Song Moon Jung, Tong Leming, Sun Ren
Department of Molecular and Medical Pharmacology, University of California at Los Angeles, Los Angeles, California 90095, USA.
J Virol. 2003 Oct;77(19):10488-503. doi: 10.1128/jvi.77.19.10488-10503.2003.
Murine gammaherpesvirus 68 (MHV-68 [also referred to as gammaHV68]) is phylogenetically related to Kaposi's sarcoma-associated herpesvirus (KSHV [also referred to as HHV-8]) and Epstein-Barr virus (EBV). However, unlike KSHV or EBV, MHV-68 readily infects fibroblast and epithelial cell lines derived from several mammalian species, providing a system to study productive and latent infections as well as reactivation of gammaherpesviruses in vivo and in vitro. To carry out rapid genome-wide analysis of MHV-68 gene expression, we made DNA arrays containing nearly all of the known and predicted open reading frames (ORFs) of the virus. RNA obtained from an MHV-68 latently infected cell line, from cells lytically infected with MHV-68 in culture, and from the lung tissue of infected mice was used to probe the MHV-68 arrays. Using a tightly latent B-cell line (S11E), the MHV-68 latent transcription program was quantitatively described. Using BHK-21 cells and infected mice, we demonstrated that latent genes are transcribed during lytic replication and are relatively independent of de novo protein synthesis. We determined that the transcription profiles at the peak of lytic gene expression are similar in cultured fibroblast and in the lung of infected mice. Finally, the MHV-68 DNA arrays were used to examine the gene expression profile of a recombinant virus that overexpresses replication and transcription activator (RTA), C-RTA/MHV-68, during lytic replication in cell culture. The recombinant virus replicates faster then the parental strain and the DNA arrays revealed that nearly every MHV-68 ORF examined was activated by RTA overexpression. Examination of the gene expression patterns of C-RTA/MHV-68 over a time course led to the finding that the M3 promoter is RTA responsive in the absence of other viral factors.
小鼠γ疱疹病毒68（MHV - 68 [也称为γHV68]）在系统发育上与卡波西肉瘤相关疱疹病毒（KSHV [也称为HHV - 8]）和爱泼斯坦 - 巴尔病毒（EBV）相关。然而，与KSHV或EBV不同，MHV - 68很容易感染源自几种哺乳动物物种的成纤维细胞和上皮细胞系，提供了一个在体内和体外研究γ疱疹病毒的生产性感染、潜伏感染以及再激活的系统。为了对MHV - 68基因表达进行快速的全基因组分析，我们制作了包含该病毒几乎所有已知和预测的开放阅读框（ORF）的DNA阵列。从MHV - 68潜伏感染的细胞系、在培养中被MHV - 68裂解感染的细胞以及感染小鼠的肺组织中获得的RNA用于探测MHV - 68阵列。使用一个紧密潜伏的B细胞系（S11E），定量描述了MHV - 68的潜伏转录程序。使用BHK - 21细胞和感染小鼠，我们证明潜伏基因在裂解复制期间被转录，并且相对独立于从头蛋白质合成。我们确定在裂解基因表达峰值时的转录谱在培养的成纤维细胞和感染小鼠的肺中相似。最后，MHV - 68 DNA阵列用于检查在细胞培养的裂解复制期间过表达复制和转录激活因子（RTA）的重组病毒C - RTA/MHV - 68的基因表达谱。重组病毒的复制速度比亲本菌株快，并且DNA阵列显示几乎每个检测的MHV - 68 ORF都被RTA过表达激活。对C - RTA/MHV - 68在一段时间内的基因表达模式进行检查发现，在没有其他病毒因子的情况下，M3启动子对RTA有反应。
