ZBP-89 regulates expression of tryptophan hydroxylase i and mucosal defense against salmonella typhimurium in mice

Bryan E. Essien, Helmut Grasberger, Rachael D. Romain, David J. Law, Natalia A. Veniaminova, Milena Saqui-Salces, Mohamad El-Zaatari, Arthur Tessier, Michael M. Hayes, Alexander C. Yang, Juanita L. Merchant

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

Background & Aims: ZBP-89 (also ZNF148 or Zfp148) is a butyrate-inducible zinc finger transcription factor that binds to GC-rich DNA elements. Deletion of the N-terminal domain is sufficient to increase mucosal susceptibility to chemical injury and inflammation. We investigated whether conditional deletion of ZBP-89 from the intestinal and colonic epithelium of mice increases their susceptibility to pathogens such as Salmonella typhimurium. Methods: We generated mice with a conditional null allele of Zfp148 (ZBP-89FL/FL) using homologous recombination to flank Zfp148 with LoxP sites (ZBP-89FL/FL), and then bred the resulting mice with those that express VillinCre. We used microarray analysis to compare gene expression patterns in colonic mucosa between ZBP-89ΔInt and C57BL/6 wild-type mice (controls). Mice were gavaged with 2 isogenic strains of S typhimurium after administration of streptomycin. Results: Microarray analysis revealed that the colonic mucosa of ZBP-89ΔInt mice had reduced levels of tryptophan hydroxylase 1 (Tph1) messenger RNA, encoding the rate-limiting enzyme in enterochromaffin cell serotonin (5-hydroxytryptamine [5HT]) biosynthesis. DNA affinity precipitation demonstrated direct binding of ZBP-89 to the mouse Tph1 promoter, which was required for its basal and butyrate-inducible expression. ZBP-89ΔInt mice did not increase mucosal levels of 5HT in response to S typhimurium infection, and succumbed to the infection 2 days before control mice. The ΔhilA isogenic mutant of S typhimurium lacks this butyrate-regulated locus and stimulated, rather than suppressed, expression of Tph1 approximately 50-fold in control, but not ZBP-89ΔInt, mice, correlating with fecal levels of butyrate. Conclusions: ZBP-89 is required for butyrate-induced expression of the Tph1 gene and subsequent production of 5HT in response to bacterial infection in mice. Reductions in epithelial ZBP-89 increase susceptibility to colitis and sepsis after infection with S typhimurium, partly because of reduced induction of 5HT production in response to butyrate and decreased secretion of antimicrobial peptides.

Original languageEnglish (US)
Pages (from-to)1466-1477.e9
JournalGastroenterology
Volume144
Issue number7
DOIs
StatePublished - Jun 2013

Bibliographical note

Funding Information:
The authors thank Lisa Travnikar for technical assistance and Laura Johnson for the SL1344 Salmonella strain. The authors acknowledge the expertise of the University of Michigan Transgenic Animal Model Core, especially Thom Saunders, Linda Samuelson and Elizabeth Hughes, supported by National Institutes of Health Cancer Center grant P30 CA46592 and Gut Peptide Center grant P30 DK34933 , and the University of Michigan Metabolic Core Service, supported by National Institutes of Health grant P30 DK089503 . The authors thank the Intrexon Corporation (Blacksburg, VA) for assembly of the pLM10-iKO-FinalTV construct and the Unit for Laboratory Animal Medicine at the University of Michigan for assistance with animal husbandry, Deborah Gumucio (University of Michigan) for the VillinCre mouse line, and Beth McCormick (University of Massachusetts, Worcester) for the hilA mutant Salmonella strain.

Keywords

  • 5HT
  • Butyrate
  • Defensin
  • ΔhilA

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