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Autographa californica multiple nucleopolyhedrovirus ac76 is involved in intranuclear microvesicle formation

Zhaoyang Hu1, Meijin Yuan, Wenbi Wu

  • 1State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China.

Journal of Virology
|May 21, 2010
PubMed

Insights

The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) orf76 (ac76) gene is essential for producing budded viruses and intranuclear microvesicles. Deleting ac76 halts viral replication, highlighting its critical role in the baculovirus life cycle.

Area of Science:

  • Virology
  • Molecular Biology
  • Insect Pathology

Background:

  • Baculoviruses, such as AcMNPV, are important pathogens of insects.
  • The function of many baculovirus genes remains unknown, including orf76 (ac76).
  • Ac76 is a highly conserved gene in lepidopteran baculoviruses.

Purpose of the Study:

  • To characterize the function of the conserved AcMNPV orf76 (ac76) gene.
  • To investigate the role of ac76 in the baculovirus life cycle.
  • To determine the involvement of ac76 in viral replication and virion formation.

Main Methods:

  • Transcriptional analysis of ac76.
  • Construction and analysis of an ac76-knockout virus.
  • Microscopy (light, electron, confocal) for viral structure and localization.
  • Titration assays and Western blot analysis for viral production.
  • Quantitative real-time PCR for viral DNA synthesis.

Main Results:

  • Ac76 transcripts initiate from a TAAG motif and have variable 3' ends.
  • The ac76-knockout virus failed to produce budded viruses.
  • Viral DNA synthesis was not affected by ac76 deletion.
  • Intranuclear microvesicles and occlusion-derived virions were absent in ac76-knockout infected cells.
  • Ac76 protein localizes to the nuclear ring zone during late infection.

Conclusions:

  • AcMNPV ac76 is essential for the production of budded viruses.
  • Ac76 plays a critical role in the formation of intranuclear microvesicles.
  • This study identifies ac76 as the first baculovirus gene involved in intranuclear microvesicle formation.

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