Genome-wide analysis reveals NRP1 as a direct HIF1α-E2F7 target in the regulation of motorneuron guidance in vivo

Nucleic Acids Res. 2016 May 5;44(8):3549-66. doi: 10.1093/nar/gkv1471. Epub 2015 Dec 17.

Abstract

In this study, we explored the existence of a transcriptional network co-regulated by E2F7 and HIF1α, as we show that expression of E2F7, like HIF1α, is induced in hypoxia, and because of the previously reported ability of E2F7 to interact with HIF1α. Our genome-wide analysis uncovers a transcriptional network that is directly controlled by HIF1α and E2F7, and demonstrates both stimulatory and repressive functions of the HIF1α -E2F7 complex. Among this network we reveal Neuropilin 1 (NRP1) as a HIF1α-E2F7 repressed gene. By performing in vitro and in vivo reporter assays we demonstrate that the HIF1α-E2F7 mediated NRP1 repression depends on a 41 base pairs 'E2F-binding site hub', providing a molecular mechanism for a previously unanticipated role for HIF1α in transcriptional repression. To explore the biological significance of this regulation we performed in situ hybridizations and observed enhanced nrp1a expression in spinal motorneurons (MN) of zebrafish embryos, upon morpholino-inhibition of e2f7/8 or hif1α Consistent with the chemo-repellent role of nrp1a, morpholino-inhibition of e2f7/8 or hif1α caused MN truncations, which was rescued in TALEN-induced nrp1a(hu10012) mutants, and phenocopied in e2f7/8 mutant zebrafish. Therefore, we conclude that repression of NRP1 by the HIF1α-E2F7 complex regulates MN axon guidance in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axon Guidance / genetics*
  • Binding Sites
  • Cell Hypoxia / genetics
  • Cell Line, Tumor
  • E2F7 Transcription Factor / genetics*
  • E2F7 Transcription Factor / metabolism
  • Genome-Wide Association Study
  • HeLa Cells
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • In Situ Hybridization
  • Morpholinos / genetics
  • Motor Neurons / metabolism*
  • Neuropilin-1 / genetics*
  • Neuropilin-1 / metabolism
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Transcription, Genetic / genetics
  • Zebrafish / embryology
  • Zebrafish / genetics*

Substances

  • E2F7 Transcription Factor
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Morpholinos
  • RNA, Small Interfering
  • Neuropilin-1