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Sequence and structural requirements of a herpes simplex viral DNA replication origin
1Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Molecular and Cellular Biology
|October 1, 1988
Summary
Researchers identified the core 75 base pair replication origin (oriS) in herpes simplex virus type 2 DNA. Essential sequences within this origin, including an AT-rich region, are crucial for viral replication initiation.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Herpes simplex virus (HSV) types 1 and 2 utilize two replication origins, oriS and oriL.
- Understanding the precise DNA sequences and structures involved in HSV replication initiation is crucial for antiviral strategies.
Purpose of the Study:
- To define the minimal DNA sequence required for the oriS replication origin of HSV type 2.
- To investigate the role of specific sequences and conformations within the oriS origin in viral DNA replication.
Main Methods:
- Construction and analysis of a series of plasmids with altered HSV type 2 oriS replication origins.
- Utilizing an in vivo replicon assay to assess the replication efficiency of modified oriS sequences.
Main Results:
- A core oriS origin of 75 base pairs (bp) was defined, encompassing a 56-bp palindrome and adjacent sequences.
- An AT-rich sequence at the center of the palindrome was found to be essential for origin function.
- Replication efficiency was enhanced by sequences flanking the core origin, and introducing additional AT-dinucleotides caused oscillating effects on origin function.
Conclusions:
- The oriS replication origin of HSV type 2 has a defined core sequence essential for initiation.
- The data suggest a linear conformation of the oriS palindrome is critical for HSV replication initiation, rather than a cruciform structure.