Open AccessCharacterization of the Cryptic AV3 Promoter of Ageratum Yellow Vein Virus in Prokaryotic and Eukaryotic Systems
Author(s) -
Wang WeiChen,
ChiaYing Wu,
Yi-Chin Lai,
NaSheng Lin,
Yau-Heiu Hsu,
Chung-Chi Hu
Publication year - 2014
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0108608
Subject(s) - nicotiana benthamiana , biology , promoter , open reading frame , transcription (linguistics) , begomovirus , escherichia coli , microbiology and biotechnology , gene , genetics , gene expression , peptide sequence , genome , linguistics , philosophy
A cryptic prokaryotic promoter, designated AV3 promoter, has been previously identified in certain begomovirus genus, including ageratum yellow vein virus isolate NT (AYVV-NT). In this study, we demonstrated that the core nucleotides in the putative −10 and −35 boxes are necessary but not sufficient for promoter activity in Escherichia coli , and showed that AYVV-NT AV3 promoter could specifically interact with single-stranded DNA-binding protein and sigma 70 of E. coli involved in transcription. Several AYVV-NT-encoded proteins were found to increase the activity of AV3 promoter. The transcription start sites downstream to AV3 promoter were mapped to nucleotide positions 803 or 805 in E. coli , and 856 in Nicotiana benthamiana . The eukaryotic activity of AV3 promoter and the translatability of a short downstream open reading frame were further confirmed by using a green fluorescent protein reporter construct in yeast ( Saccharomyces cerevisiae ) cells. These results suggested that AV3 promoter might be a remnant of evolution that retained cryptic activity at present.