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Mutational analysis of the herpes simplex virus type 1 trans-inducing factor Vmw65
C I Ace1, M A Dalrymple, F H Ramsay
1Medical Research Council Virology Unit, Glasgow, U.K.
The Journal of General Virology
|October 1, 1988
Summary
Herpes simplex virus type 1 (HSV-1) Vmw65 protein activates immediate early (IE) transcription by forming a protein-DNA complex (IEC). Distinct regions of Vmw65 are essential for both transcription activation and virus assembly.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Herpes simplex virus type 1 (HSV-1) Vmw65 is crucial for viral particle structure and immediate early (IE) transcription.
- Understanding Vmw65's functional domains is key to deciphering HSV-1 replication mechanisms.
Purpose of the Study:
- To investigate the functional domains of the HSV-1 Vmw65 polypeptide.
- To correlate Vmw65's role in IE transcription with its function in virus assembly.
Main Methods:
- Constructed 10 plasmids with 12 bp insertions in the Vmw65 gene.
- Assayed plasmid ability to stimulate IE transcription via transfection.
- Tested Vmw65 variants for IE-specific protein-DNA complex (IEC) formation in vitro.
- Evaluated plasmid ability to rescue HSV-2 ts2203 assembly mutant.
Main Results:
- A direct correlation was found between IE transcription stimulation and IEC formation, indicating IEC's role in activation.
- Five plasmids failed to rescue the ts2203 mutant, identifying Vmw65 regions essential for virus assembly.
- Distinct functional domains within Vmw65 were identified for transcription activation and virus assembly.
Conclusions:
- The IE-specific protein-DNA complex (IEC) is a critical intermediate for Vmw65-mediated transcription activation.
- Vmw65 possesses separate domains responsible for activating transcription and facilitating virus assembly.
- These findings provide insights into the multifunctional nature of Vmw65 in the HSV-1 life cycle.