The Chilo iridescent virus DNA polymerase promoter contains an essential AAAAT motif


Nalcacioglu R. , İNCE I. A. , VLAK J. M. , Demirbag Z. , van Oers M. M.

JOURNAL OF GENERAL VIROLOGY, cilt.88, ss.2488-2494, 2007 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 88
  • Basım Tarihi: 2007
  • Doi Numarası: 10.1099/vir.0.82947-0
  • Dergi Adı: JOURNAL OF GENERAL VIROLOGY
  • Sayfa Sayıları: ss.2488-2494

Özet

The delayed-early DNA polymerase promoter of Chilo iridescent virus (CIV), officially known as Invertebrate iridescent virus, was fine mapped by constructing a series of increasing deletions and by introducing point mutations. The effects of these mutations were examined in a luciferase reporter gene system using Bombyx mori cells transfected with promoter constructs and infected with CIV. When the size of the upstream element was reduced from position -19 to -15, relative to the transcriptional start site, the luciferase activity was reduced to almost zero. Point mutations showed that each of the 5 nt (AAAAT) located between -19 and -15 were equally essential for promoter activity. Mutations at individual bases around the transcription initiation site showed that the promoter extended until position -2 upstream of the transcription start site. South-Western analysis showed that a protein of approximately 100 kDa interacted with the -19 nt promoter fragment in CIV-infected cells. This binding did not occur with a point mutant that lacked promoter activity. The AAAAT motif was also found in the DNA polymerase promoter region of other iridoviruses and in other putative CIV delayed-early genes.