RRC ID 41730
Author Hirata Y, Maeda S, Mitsuno Y, Tateishi K, Yanai A, Akanuma M, Yoshida H, Kawabe T, Shiratori Y, Omata M.
Title Helicobacter pylori CagA protein activates serum response element-driven transcription independently of tyrosine phosphorylation.
Journal Gastroenterology
Abstract BACKGROUND & AIMS:Infection with Helicobacter pylori possessing the cag pathogenicity island (PAI) is associated with severe gastritis and gastric cancer. CagA protein, one of the products of cag PAI, is translocated into epithelial cells, where cytoskeletal rearrangements occur as a result of CagA tyrosine (Tyr) phosphorylation. Here we identify a new role for CagA protein as an activator of host cell signaling.
METHODS:We transfected CagA into epithelial cells and analyzed its effect on transcription by reporter assays. The mechanism of reporter activation was assessed by electrophoretic mobility shift assays (EMSA) and immunoblots. Responsible regions of CagA for reporter activation were determined by truncation and mutagenesis of cagA gene.
RESULTS:In HeLa cells, expression of CagA increased serum response element (SRE)-driven and serum response factor (SRF)-driven transcription by 40-fold and 3.3-fold, respectively, but did not affect nuclear factor kappaB- or AP-1-driven transcription. CagA-mediated SRE activation was also observed in gastric cell lines. Immunoblotting and EMSA revealed that transfection of CagA enhanced phosphorylation of and DNA binding by Elk1. Furthermore, involvement of Ras and MEK in CagA-mediated Elk1 phosphorylation was observed. SRE activation was dependent on several regions within the C-terminal portion of CagA (CagA(873-1002)), and independent of Tyr phosphorylation.
CONCLUSIONS:The C-terminal portion of CagA enhances SRE-driven transcription by activating an upstream signaling cascade without requiring CagA Tyr phosphorylation. This result suggests that translocated CagA regulates 2 distinct cellular responses: phosphorylation-dependent cytoskeletal rearrangement and phosphorylation-independent transcriptional activation.
Volume 123(6)
Pages 1962-71
Published 2002-12-1
DOI 10.1053/gast.2002.37044
PII S0016508502004687
PMID 12454853
IF 17.373
Times Cited 66
Human and Animal Cells HeLa(RCB0007)