Mechanism(S) Involved in the Colon-Specific Expression of the Thiamine Pyrophosphate (Tpp) Transporter

• Published in: PloS one 2016-02, Vol.11 (2), p.e0149255
• Main Authors: Nabokina, Svetlana M, Ramos, Mel Brendan, Said, Hamid M
• Format: Article
• Language: English
• Subjects: 3' Untranslated Regions; Animals; Antibiotics; Azacitidine - pharmacology; Base Sequence; Biochemistry; Biological Transport; Biology; Biology and life sciences; Biophysics; Cell Line; Cloning; Colon; Colon - metabolism; CpG Islands; Deoxyribonucleic acid; DNA; DNA Methylation; Epigenetics; Epithelial cells; Gastroenterology; Gastrointestinal tract; Gene expression; Gene Expression Regulation - drug effects; Genes; Genomes; Histone H3; Humans; Jejunum; Large intestine; Lysine; Medicine and Health Sciences; Membrane Transport Proteins - genetics; Metabolism; Mice; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Molecular Sequence Data; Organ Specificity - genetics; Physiology; Proteins; Retina; Ribonucleic acid; RNA; RNA-binding protein; RNA-Binding Proteins; Rodents; Sequence Alignment; Statistical analysis; Target recognition; Thiamine; Thiamine Pyrophosphate - metabolism; Trans-Pacific Partnership Agreement; Transcription factors; Vitamin B; Vitamin B1
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0149255

Microbiota of the large intestine synthesizes considerable amount of vitamin B1 (thiamine) in the form of thiamine pyrophosphate (TPP). We have recently demonstrated the existence of an efficient and specific carrier-mediated uptake process for TPP in human colonocytes, identified the TPP transporter (TPPT) involved (product of the SLC44A4 gene), and shown that expression of TPPT along the gastrointestinal (GI) tract is restricted to the colon. Our aim in this study was to determine the molecular basis of the colon-specific expression of TPPT focusing on a possible epigenetic mechanism. Our results showed that the CpG island predicted in the SLC44A4 promoter is non-methylated in the human colonic epithelial NCM460 cells, but is hyper-methylated in the human duodenal epithelial HuTu80 cells (as well as in the human retinal pigment epithelial ARPE19 cells). In the mouse (where TPPT expression in the GI tract is also restricted to the colon), the CpG island predicted in the Slc44a4 promoter is non-methylated in both the jejunum and colon, thus arguing against possible contribution of DNA methylation in the colon-specific expression of TPPT. A role for histone modifications in the tissue-specific pattern of Slc44a4 expression, however, was suggested by the findings that in mouse colon, histone H3 in the 5'-regulatory region of Slc44a4 is tri-methylated at lysine 4 and acetylated at lysine 9, whereas the tri-methylation at lysine 27 modification was negligible. In contrast, in the mouse jejunum, histone H3 is hyper-trimethylated at lysine 27 (repressor mark). Similarly, possible involvement of miRNA(s) in the tissue-specific expression of TPPT was also suggested by the findings that the 3'-UTR of SLC44A4 is targeted by specific miRNAs/RNA binding proteins in non-colonic, but not in colonic, epithelial cells. These studies show, for the first time, epigenetic mechanisms (histone modifications) play a role in determining the tissue-specific pattern of expression of TPPT in the GI tract.