title: The tripartite leader sequence is required for ectopic expression of HAdV-B and HAdV-E E3 CR1 genes.
abstract: The unique repertoire of genes that characterizes the early region 3 (E3) of the different species of human adenovirus (HAdV) likely contributes to their distinct pathogenic traits. The function of many E3 CR1 proteins remains unknown possibly due to unidentified intrinsic properties that make them difficult to express ectopically. This study shows that the species HAdV-B- and HAdV-E-specific E3 CR1 genes can be expressed from vectors carrying the HAdV tripartite leader (TPL) sequence but not from traditional mammalian expression vectors. Insertion of the TPL sequence upstream of the HAdV-B and HAdV-E E3 CR1 open reading frames was sufficient to rescue protein expression from pCI-neo constructs in transfected 293T cells. The detection of higher levels of HAdV-B and HAdV-E E3 CR1 transcripts suggests that the TPL sequence may enhance gene expression at both the transcriptional and translational levels. Our findings will facilitate the characterization of additional AdV E3 proteins.
authors.affiliation: Infectious Disease Program, Lovelace Respiratory Research Institute, Albuquerque, NM, United States.
authors.forename: Camden R
authors.initials: CR
authors.lastname: Bair
authors.affiliation: Infectious Disease Program, Lovelace Respiratory Research Institute, Albuquerque, NM, United States.
authors.forename: Poornima
authors.initials: P
authors.lastname: Kotha Lakshmi Narayan
authors.affiliation: Infectious Disease Program, Lovelace Respiratory Research Institute, Albuquerque, NM, United States. Electronic address: akajon@lrri.org.
authors.forename: Adriana E
authors.initials: AE
authors.lastname: Kajon
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics*; D017503:Adenovirus E3 Proteins / Q000096:biosynthesis* / Q000235:genetics*; D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics*; D000066630:Ectopic Gene Expression / Q000235:genetics*; D018274:Electroporation; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D016366:Open Reading Frames / Q000235:genetics; D021382:Protein Sorting Signals; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; D017503:Adenovirus E3 Proteins; D021382:Protein Sorting Signals; D014764:Viral Proteins
keywords: CR1 genes; Early region 3; Human adenovirus; Tripartite leader sequence; pMT2