Key elements of the human bocavirus type 1 HBoV1 promoter and its trans-activation by NS1 proteinReportar como inadecuado




Key elements of the human bocavirus type 1 HBoV1 promoter and its trans-activation by NS1 protein - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Virology Journal

, 10:315

First Online: 27 October 2013Received: 05 February 2013Accepted: 23 October 2013

Abstract

BackgroundHuman bocavirus HBoV, a parvovirus, is suspected to be an etiologic agent of respiratory disease and gastrointestinal disease in humans. All mRNAs of HBoV1 are transcribed from a single promoter.

MethodsIn this study, we constructed EGFP and luciferase reporter gene vectors under the control of the HBoV1 full promoter nt 1–252 and its mutated variants, respectively. Fluorescence microscopy was used to observe expression activities of the EGFP. Dual-luciferase reporter vectors were employed in order to evaluate critical promoter elements and the effect of NS1 protein on promoter activity.

ResultsThe HBoV1 promoter activity was about 2.2-fold and 1.9-fold higher than that of the CMV promoter in 293 T and HeLa cells, respectively. The putative transcription factor binding region of the promoter was identified to be located between nt 96 and nt 145. Mutations introduced in the CAAT box of the HBoV1 promoter reduced promoter activity by 34%, whereas nucleotide substitutions in the TATA box had no effect on promoter activity. The HBoV1 promoter activities in 293 T and HeLa cells, in the presence of NS1 protein, were 2- to 2.5-fold higher than those in the absence of NS1 protein.

ConclusionThe HBoV1 promoter was highly active in 293 T and HeLa cell lines, and the sequence from nt 96 to nt 145 was critical for the activity of HBoV1 promoter. The CAAT box, in contrast to the TATA-box, was important for optimum promoter activity. In addition, the transcriptional activity of this promoter could be trans-activated by the viral nonstructural protein NS1 in these cells.

KeywordsHBoV1 Promoter activity NS1 protein Trans-activation Electronic supplementary materialThe online version of this article doi:10.1186-1743-422X-10-315 contains supplementary material, which is available to authorized users.

Download fulltext PDF



Autor: Jingjing Li - Yongbo Yang - Yanming Dong - Yongshu Li - Yu Huang - Qianhui Yi - Kaiyu Liu - Yi Li

Fuente: https://link.springer.com/







Documentos relacionados