A Structural Investigation into Oct4 Regulation by Orphan Nuclear Receptors, Germ Cell Nuclear Factor (GCNF), and Liver Receptor Homolog-1 (LRH-1)

J Mol Biol. 2016 Dec 4;428(24 Pt B):4981-4992. doi: 10.1016/j.jmb.2016.10.025. Epub 2016 Oct 27.

Abstract

Oct4 is a transcription factor required for maintaining pluripotency and self-renewal in stem cells. Prior to differentiation, Oct4 must be silenced to allow for the development of the three germ layers in the developing embryo. This fine-tuning is controlled by the nuclear receptors (NRs), liver receptor homolog-1 (LRH-1) and germ cell nuclear factor (GCNF). Liver receptor homolog-1 is responsible for driving the expression of Oct4 where GCNF represses its expression upon differentiation. Both receptors bind to a DR0 motif located within the Oct4 promoter. Here, we present the first structure of mouse GCNF DNA-binding domain in complex with the Oct4 DR0. The overall structure revealed two molecules bound in a head-to-tail fashion on opposite sides of the DNA. Additionally, we solved the structure of the human LRH-1 DNA-binding domain bound to the same element. We explore the structural elements that govern Oct4 recognition by these two NRs.

Keywords: Oct4 regulation; X-ray crystallography; germ cell nuclear factor; liver receptor homolog-1; nuclear receptors.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Crystallography, X-Ray
  • DNA / metabolism
  • Gene Expression Regulation, Developmental
  • Humans
  • Mice
  • Models, Molecular
  • Nuclear Receptor Subfamily 6, Group A, Member 1 / chemistry*
  • Nuclear Receptor Subfamily 6, Group A, Member 1 / metabolism*
  • Octamer Transcription Factor-3 / biosynthesis*
  • Octamer Transcription Factor-3 / chemistry*
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Conformation
  • Receptors, Cytoplasmic and Nuclear / chemistry*
  • Receptors, Cytoplasmic and Nuclear / metabolism*

Substances

  • NR5A2 protein, human
  • Nr6a1 protein, mouse
  • Nuclear Receptor Subfamily 6, Group A, Member 1
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse
  • Receptors, Cytoplasmic and Nuclear
  • DNA