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Guinea pig cytomegalovirus immediate-early transcription
1Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey 17033.
Abstract:
Guinea pig cytomegalovirus (GPCMV) immediate-early (IE) gene expression was analyzed. GPCMV IE RNA was defined as RNA obtained from GPCMV-infected guinea pig cells treated with cycloheximide for 1 h before infection and for 4 h postinfection. Mapping studies showed that GPCMV IE genes are located at several distinct sites on the GPCMV genome. A total of 17 GPCMV IE transcripts were identified, and 9 IE transcripts coded for by three specific regions of the genome (regions I, II, and III) were characterized in detail. A series of recombinant DNA clones were generated to identify the nine IE transcripts. Three of the IE transcripts from region I and three from region III were transcribed in the same direction from overlapping sequences. The 2.0-kilobase (kb) transcript encoded by the EcoRI E DNA fragment (region II) was the most abundant IE GPCMV transcript. The cloned GPCMV DNA subfragment that was used to identify the region II EcoRI E 2.0-kb transcript did not hybridize to GPCMV early or late RNA, indicating that this transcript is expressed only under IE conditions. Expression of RNAs from the IE genes was also measured during a natural GPCMV infection in the absence of cycloheximide. During the natural infection, the transcripts previously identified under IE cycloheximide block conditions were expressed, and the region II EcoRI E 2.0-kb transcript was the most abundant transcript at 1 h postinfection. In addition, a rise and fall in RNA levels was observed during the natural infection, demonstrating the transient nature of expression of these transcripts. We conclude that GPCMV IE gene expression is complex, involving a reasonably large number of genes, and demonstrates some similarities with IE transcription by other CMVs.
Insights
Guinea pig cytomegalovirus (GPCMV) immediate-early (IE) gene expression involves multiple transcripts and regions. The study identified 17 IE transcripts, with a 2.0-kb transcript being the most abundant during natural GPCMV infection.
Area of Science:
- Virology
- Molecular Biology
- Genomics
Background:
- Cytomegaloviruses (CMVs) are significant human pathogens.
- Understanding immediate-early (IE) gene expression is crucial for viral replication.
- Guinea pig cytomegalovirus (GPCMV) serves as a model for CMV research.
Purpose of the Study:
- To analyze the immediate-early (IE) gene expression of Guinea pig cytomegalovirus (GPCMV).
- To identify and characterize GPCMV IE transcripts and their genomic locations.
- To compare GPCMV IE gene expression with other CMV species.
Main Methods:
- GPCMV-infected guinea pig cells were treated with cycloheximide to isolate IE RNA.
- RNA mapping and Northern blot hybridization were used to identify and characterize IE transcripts.
- Recombinant DNA clones were generated to pinpoint specific IE transcripts.
Main Results:
- Seventeen GPCMV IE transcripts were identified, with nine characterized from specific genomic regions (I, II, and III).
- A 2.0-kilobase (kb) transcript from the EcoRI E DNA fragment (region II) was the most abundant IE transcript.
- This 2.0-kb transcript was exclusively expressed under IE conditions and was the most abundant at 1 hour postinfection during natural GPCMV infection.
Conclusions:
- GPCMV IE gene expression is complex, involving a substantial number of genes.
- The identified IE transcripts show transient expression patterns during natural infection.
- GPCMV IE transcription shares similarities with that of other characterized cytomegaloviruses.