hSNF5 is required for human papillomavirus E2-driven transcriptional activation and DNA replication.
OBJECTIVE
To determine the participation of the SWI/SNF complex in the transcription and replication of human papillomavirus (HPV) E2 protein.


METHODS
We checked the interaction between hSNF5 and HPV E2 through glutathione S-transferase (GST) pull-down and coimmunoprecipitation assays. The transcriptional activation of E2 was analyzed by reporter assay and the level of HPV DNA replication was determined by a transient DNA replication assay.


RESULTS
We demonstrated that hSNF5 binds to the HPV E2 protein in vivo and in vitro and activates E2-dependent viral transcription. SWI/SNF components enhanced E2-dependent viral transcription. The ATPase activity of BRG-1/hSNF2 was required for efficient E2-dependent transcriptional activation. Transient DNA replication assays showed that hSNF5 and BRG-1 enhance HPV-18 DNA replication in vivo. A dominant negative hSNF5 and a BRG-1 ATPase mutant each repressed E2-dependent viral transcription and E2-driven HPV DNA replication in vivo. The fact that the transcriptional activation function of HPV-18 E2 was defective in the SNF5 knockout strains of the yeast Saccharomyces cerevisiae implies that the SWI/SNF complex is required for the transcriptional activation function of E2.


CONCLUSIONS
These results suggest that the SWI/SNF complex is involved in HPV E2-driven transcription and DNA replication via interaction with E2.
Cha
Seho
S
Department of Life Science, Dongguk University-Seoul, Seoul, South Korea.
Seo
Taegun
T
D002460:Cell Line; D002868:Chromosomal Proteins, Non-Histone; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D027383:Papillomaviridae / Q000472:pathogenicity*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D000071796:SMARCB1 Protein; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014157:Transcription Factors; D015533:Transcriptional Activation
D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D009856:Oncogene Proteins, Viral; D000071796:SMARCB1 Protein; C513266:SMARCB1 protein, human; D014157:Transcription Factors; C069102:oncogene protein E2, Human papillomavirus type 18