IL-10 promoter transactivation by the viral K-RTA protein involves the host-cell transcription factors, specificity proteins 1 and 3

J Biol Chem. 2018 Jan 12;293(2):662-676. doi: 10.1074/jbc.M117.802900. Epub 2017 Nov 28.

Abstract

Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus-8 (HHV-8) causes a persistent infection, presenting latent and lytic replication phases during its life cycle. KSHV-related diseases are associated with deregulated expression of inflammatory cytokines, including IL-6 and IL-10, but the mechanisms underlying this dysregulation are unclear. Herein, we report a molecular mechanism for KSHV-induced IL-10 gene expression. KSHV replication and transcription activator (K-RTA) is a molecular switch for the initiation of expression of viral lytic genes, and we describe, for the first time, that K-RTA significantly activates the promoter of the human IL-10 gene. Of note, mutations involving a basic region of K-RTA reduced the association of K-RTA with the IL-10 promoter. Moreover, the host-cell transcription factors, specificity proteins (SP) 1 and 3, play a pivotal cooperative role in K-RTA-mediated transactivation of the IL-10 promoter. K-RTA can interact with SP1 and SP3 directly in vitro, and electrophoresis mobility shift assays (EMSAs) revealed co-operative interaction involving K-RTA, SP1, and SP3 in binding to the IL-10 promoter. As DNase I footprinting assays indicated that K-RTA did not affect SP3 binding to the IL-10 promoter, SP3 can function to recruit K-RTA to the IL-10 promoter. These findings indicate that K-RTA can directly contribute to IL-10 up-regulation via a functional interplay with the cellular transcription factors SP1 and SP3.

Keywords: IL-10; SP3; herpesvirus; interleukin; specificity protein 1 (Sp1); transactivation; transcription; viral transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites / genetics
  • Binding Sites / physiology
  • Gene Expression Regulation, Viral / genetics
  • Gene Expression Regulation, Viral / physiology
  • Humans
  • Interleukin-10 / genetics*
  • Interleukin-10 / metabolism*
  • Promoter Regions, Genetic / genetics
  • Sp1 Transcription Factor / genetics
  • Sp1 Transcription Factor / metabolism
  • Sp3 Transcription Factor / genetics
  • Sp3 Transcription Factor / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcriptional Activation / genetics
  • Transcriptional Activation / physiology
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*
  • Virus Replication / genetics
  • Virus Replication / physiology

Substances

  • IL10 protein, human
  • SP3 protein, human
  • Sp1 Transcription Factor
  • SP1 protein, human
  • Transcription Factors
  • Viral Proteins
  • Interleukin-10
  • Sp3 Transcription Factor