Mapping of the Sequences Directing Localization of the Drosophila Germ Cell-Expressed Protein (GCE)

PLoS One. 2015 Jul 17;10(7):e0133307. doi: 10.1371/journal.pone.0133307. eCollection 2015.

Abstract

Drosophila melanogaster germ cell-expressed protein (GCE) belongs to the family of bHLH-PAS transcription factors that are the regulators of gene expression networks that determine many physiological and developmental processes. GCE is a homolog of D. melanogaster methoprene tolerant protein (MET), a key mediator of anti-metamorphic signaling in insects and the putative juvenile hormone receptor. Recently, it has been shown that the functions of MET and GCE are only partially redundant and tissue specific. The ability of bHLH-PAS proteins to fulfill their function depends on proper intracellular trafficking, determined by specific sequences, i.e. the nuclear localization signal (NLS) and the nuclear export signal (NES). Nevertheless, until now no data has been published on the GCE intracellular shuttling and localization signals. We performed confocal microscopy analysis of the subcellular distribution of GCE fused with yellow fluorescent protein (YFP) and YFP-GCE derivatives which allowed us to characterize the details of the subcellular traffic of this protein. We demonstrate that GCE possess specific pattern of localization signals, only partially consistent with presented previously for MET. The presence of a strong NLS in the C-terminal part of GCE, seems to be unique and important feature of this protein. The intracellular localization of GCE appears to be determined by the NLSs localized in PAS-B domain and C-terminal fragment of GCE, and NESs localized in PAS-A, PAS-B domains and C-terminal fragment of GCE. NLSs activity can be modified by juvenile hormone (JH) and other partners, likely 14-3-3 proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Chlorocebus aethiops
  • Drosophila Proteins / chemistry*
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / drug effects
  • Drosophila melanogaster / metabolism*
  • HEK293 Cells
  • Humans
  • Intracellular Space / metabolism
  • Juvenile Hormones / pharmacology
  • Molecular Sequence Data
  • Mutant Proteins / chemistry
  • Mutant Proteins / metabolism
  • Nuclear Export Signals
  • Nuclear Localization Signals / chemistry
  • Nuclear Localization Signals / metabolism
  • Protein Structure, Tertiary
  • Protein Transport / drug effects
  • Sequence Analysis, Protein*
  • Subcellular Fractions / drug effects
  • Subcellular Fractions / metabolism
  • Transcription Factors / chemistry*
  • Transcription Factors / metabolism*

Substances

  • Drosophila Proteins
  • Juvenile Hormones
  • Mutant Proteins
  • Nuclear Export Signals
  • Nuclear Localization Signals
  • Transcription Factors
  • gce protein, Drosophila

Grants and funding

This work has been supported by a statutory activity subsidy from The Polish Ministry of Science and High Education for the Faculty of Chemistry of Wrocław University of Technology. Publication supported by Wroclaw Centre of Biotechnology, programme The Leading National Research Centre (KNOW) for years 2014-2018.