Tiled microarray identification of novel viral transcript structures and distinct transcriptional profiles during two modes of productive murine gammaherpesvirus 68 infection.
We applied a custom tiled microarray to examine murine gammaherpesvirus 68 (MHV68) polyadenylated transcript expression in a time course of de novo infection of fibroblast cells and following phorbol ester-mediated reactivation from a latently infected B cell line. During de novo infection, all open reading frames (ORFs) were transcribed and clustered into four major temporal groups that were overlapping yet distinct from clusters based on the phorbol ester-stimulated B cell reactivation time course. High-density transcript analysis at 2-h intervals during de novo infection mapped gene boundaries with a 20-nucleotide resolution, including a previously undefined ORF73 transcript and the MHV68 ORF63 homolog of Kaposi's sarcoma-associated herpesvirus vNLRP1. ORF6 transcript initiation was mapped by tiled array and confirmed by 5' rapid amplification of cDNA ends. The ∼1.3-kb region upstream of ORF6 was responsive to lytic infection and MHV68 RTA, identifying a novel RTA-responsive promoter. Transcription in intergenic regions consistent with the previously defined expressed genomic regions was detected during both types of productive infection. We conclude that the MHV68 transcriptome is dynamic and distinct during de novo fibroblast infection and upon phorbol ester-stimulated B cell reactivation, highlighting the need to evaluate further transcript structure and the context-dependent molecular events that govern viral gene expression during chronic infection.
Department of Molecular Genetics and Microbiology, State University of New York—Stony Brook, Stony Brook, New York, USA.
Benson Yee Hin
BY
Cheng
Jizu
J
Zhi
Alexis
A
Santana
Sohail
S
Khan
Eduardo
E
Salinas
J Craig
JC
Forrest
Yueting
Y
Zheng
Shirin
S
Jaggi
Janet
J
Leatherwood
Laurie T
LT
Krug
D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D002460:Cell Line; D016000:Cluster Analysis; D019295:Computational Biology; D005347:Fibroblasts / Q000378:metabolism / Q000821:virology; D018142:Gammaherpesvirinae / Q000235:genetics*; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D016366:Open Reading Frames; D050436:Regulatory Elements, Transcriptional; D015203:Reproducibility of Results; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D059467:Transcriptome
D013755:Tetradecanoylphorbol Acetate