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RNA polymerase domain of NS5 protein from Yellow Fever virus interacts with nuclear cellular proteins
Author(s) -
Nogueira Mauricio Lacerda,
Madrid Maria C F S,
Bronzoni Roberta V M,
Carmo Ana C V,
Zanelli Cleslei,
Valentini Sandro R
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.lb30-d
Subject(s) - biology , microbiology and biotechnology , rna polymerase , polymerase , rna splicing , gene , transcription (linguistics) , nuclear localization sequence , rna , virology , genetics , linguistics , philosophy
Yellow Fever virus (YFV) is a member of the flaviviridae family. NS5 is a viral dual function protein that has a methyltransferase and a RNA polymerase domain, and despite its cytoplasmic function is also detected in the nucleus of infected cells. However, its nuclear function is not known. The aim of this study was to identify and characterize the interactions of NS5 protein with cellular proteins. YFV NS5 RNA polymerase fragment of NS5 gene was cloned in a DNA‐BD plasmid. For the two‐hybrid system screening, RNApol bait and the prey, (AD vector/human cDNA library), were transformed into AH109 strain of Saccharomyces cerevisiae . The transformants, initially selected in SD‐HIS‐LEU‐TRP drop‐out media, were tested for the activation of three AH109 yeast reporters’ genes. Ten out of 100 positive colonies for HIS3, ADE2 and lacZ reporters were selected for further studies. These clones included protein involved in RNA splicing and translation, transcription activation and regulation, apoptosis control, and cytokines. Deletions in NS5 showed that these interactions use different parts of the protein. These data suggests that NS5 is a multifunctional protein and is related to several activities including actions in the nucleus of the cell. Financial Support: (FAPESP)