Characterization of the Role of individual Protein Binding Motifs within the Hepatitis B Virus Enhancer I on X Promoter Activity Using Linker Scanning Mutagenesis

Kurt E Gustin, Mark Shapiro, Wendy Lee, Robert D. Burk

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

A combination of linker scanning mutagenesis and deletional analyses has been used to determine the role of individual DNA:protein binding sites on expression from the hepatitis B virus (HBV) enhancer I-X promoter (map position (mp) 1042-1354, HBV adw2). Linker scanning mutation of the EF-C site caused a 67.5% drop in X promoter activity in HuH7 cells, but had no effect in HepG2 or HepSK cells. Mutation of the E element resulted in an approximately 50% reduction in X promoter activity in HUH7. HepG2, and HepSK cells. Deletional analysis showed that sequences upstream of the EF-C site (mp 1163) were required for full X promoter activity and implicated the NF-1 a site as being sufficient for basal X promoter activity. However, PCR-directed linker scanning mutation of the NF-1 a site did not cause a reduction in X promoter activity, indicating that this site was not an essential component of the X promoter. Taken together, these results indicated that multiple, partially redundant protein:DNA interactions in the enhancer I are essential for full X promoter activity. The lack of an essential basal promoter element supports the suggestion that the two separate HBV enhancer elements (enhI and enhII) were created by integration of the X gene into a primordial enhancer element.

Original languageEnglish (US)
Pages (from-to)653-660
Number of pages8
JournalVirology
Volume193
Issue number2
DOIs
StatePublished - Apr 1993
Externally publishedYes

Fingerprint

Amino Acid Motifs
Hepatitis B virus
Protein Binding
Mutagenesis
Mutation
DNA
Hep G2 Cells
Sequence Analysis
Proteins
Binding Sites
Polymerase Chain Reaction
Genes

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

Cite this

Characterization of the Role of individual Protein Binding Motifs within the Hepatitis B Virus Enhancer I on X Promoter Activity Using Linker Scanning Mutagenesis. / Gustin, Kurt E; Shapiro, Mark; Lee, Wendy; Burk, Robert D.

In: Virology, Vol. 193, No. 2, 04.1993, p. 653-660.

Research output: Contribution to journalArticle

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