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URL: https://dx.doi.org/DOI:10.17504/protocols.io.pnidmce

Authors: Kaur G, Helmer RA, Smith LA, Martinez-Zaguilan R, Dufour JM, Chilton BS

Summary: Helicase-like transcription factor is regulated by alternative mRNA splicing. Global deletion of Hltf causes perinatal lethality, i.e. 75% die of hypoglycemia Hif-1α that regulates transport systems. Thus, we tested the hypothesis that Hltf deletion in placenta either caused or exacerbated neonatal hypoglycemia via Hif-1α regulation of nutrient transporters.  RNA-seq data analyses of the placentome identified significant changes in transcript expression and splicing. The iPathwayGuide tool was used for gene ontology (GO) analysis of biological processes, molecular functions and cellular components. The Elim pruning algorithm identified hierarchical relationships. The methylome was evaluated by Methyl-MiniSeq Epiquest analysis. GO analysis with DAVID identified gene enrichment within biological processes. Protein expression was visualized with immunohistochemistry. Although two Hltf mRNA isoforms are quantifiable in most murine tissues, only the truncated Hltf isoform is expressed in placenta. iPathwayGuide analysis identified targets − 157 genes of 11,538 total genes with measured expression – of the encoded Hltf protein. Hltf deletion altered transcription of trophoblast lineage-specific genes, and increased transcription of the Cxcr7 (p=0.004) gene whose protein product is a co-receptor for human and simian immunodeficiency viruses. Concomitant increased Cxcr7 protein was identified with immune labeling. Hltf gene deletion had no effect on transcription or site-specific methylation patters of Hif-1α, the major glucose transporters, or System A amino acid transporters. There was no measureable evidence of uteroplacental dysfunction or fetal compromise. iPathGuide analysis revealed Hltf suppresses cytolysis (10/21 genes; p-value 1.900e-12; p-value correction: Elim pruning; GO:019835) including the perforin-granzyme pathway in uterine natural killer cells. Our findings 1) identify a functional link between alternative splicing of Hltf and immunosuppression at the feto-maternal interface, and 2) underscore the importance of differential splicing analysis to identify functional diversity in the transcription factor under investigation.

Associated Publications: Kaur G, Helmer RA, Smith LA, Martinez-Zaguilan R, Dufour JM, Chilton BS (2018) Alternative splicing of helicase-like transcription factor (Hltf): Intron retention-dependent activation of immune tolerance at the feto-maternal interface. PLoS ONE 13(7): e0200211. doi: 10.1371/journal.pone.0200211

Affiliations: Texas Tech University-Health Sciences Center, Texas Tech University-Health Sciences Center, Texas Tech University-Health Sciences Center, Texas Tech University-Health Sciences Center, Texas Tech University-Health Sciences Center, Texas Tech University-Health Sciences Center

External URL: https://doi.org/10.1371/journal.pone.0200211

Version: 1

Publication Date: 2018

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