Related Experiment Videos
Multicomponent origin of cytomegalovirus lytic-phase DNA replication
1Virology Laboratories, Wadsworth Center for Laboratories and Research, State University of New York, Albany.
Journal of Virology
|February 1, 1991
Summary
Researchers identified the origin of Cytomegalovirus (CMV) replication, oriLyt, essential for viral DNA synthesis. This complex region, located upstream of the DNA-binding protein gene, is crucial for initiating CMV lytic-phase replication.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Cytomegalovirus (CMV) lytic-phase DNA replication needs viral factors and a specific origin site.
- The precise location and structure of the CMV replication origin (oriLyt) remained undefined.
Purpose of the Study:
- To identify and characterize the cis-acting element, oriLyt, essential for CMV lytic-phase DNA replication.
- To investigate the structural requirements of oriLyt for its function.
Main Methods:
- Assessing cloned DNA fragments for their ability to mediate phosphonoformic acid-sensitive DNA replication in human fibroblasts.
- Utilizing transient assays and deletion analyses to define the minimal origin region.
- Nucleotide sequencing of the identified origin region.
Main Results:
- A candidate origin of replication, oriLyt, was located in both simian and human CMV strains.
- Subcloning and deletion analyses defined a 1,300 bp region upstream of the DNA-binding protein gene (dbp) as sufficient for origin function.
- The oriLyt region contains four distinct domains, including repeated sequences and an A+T-rich segment, all required for function.
Conclusions:
- CMV oriLyt is a structurally complex origin of DNA replication, similar to that of Epstein-Barr virus.
- The identified oriLyt is essential for initiating CMV lytic-phase DNA replication.