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Functional dissection of the baculovirus late expression factor-8 gene: sequence requirements for late gene promoter activation

Jane S. Titterington

Tamara K. Nun

Molecular, Cellular and Developmental Biology Program, Division of Biology, Kansas State University, 232 Ackert Hall, Manhattan, KS 66506-4901, USA

Correspondence
Lorena Passarelli
lpassar{at}ksu.edu

The late expression factor-8 gene (lef-8) of Autographa californica M nucleopolyhedrovirus encodes the largest subunit of the virally encoded DNA-directed RNA polymerase specific for the transcription of late and very late viral genes. The sequence of lef-8 predicts a C-terminal motif of 13 amino acids that is conserved in other polymerases. Detailed mutagenesis throughout lef-8 was performed, including this C-terminal motif, to define sequences required for late promoter activation. It was found that the conserved C-terminal motif was critical for late gene expression. In addition, regions throughout the entire lef-8-encoding sequence were important for optimal function, suggesting complex protein–protein and protein–DNA interrelationships in the late gene-specific viral transcriptosome.

Present address: University of Kansas Medical Center, Kansas City, KS 66160, USA.

Present address: University of North Carolina, Chapel Hill, NC 27599, USA.

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