title: Mapping of the Sequences Directing Localization of the Drosophila Germ Cell-Expressed Protein (GCE).
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.
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, Wrocław University of Technology, Wrocław, Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Greb-Markiewicz
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, Wrocław University of Technology, Wrocław, Poland.
authors.forename: Daria
authors.initials: D
authors.lastname: Sadowska
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, Wrocław University of Technology, Wrocław, Poland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Surgut
authors.affiliation: Department of Neurosurgery, Brigham and Woman's Hospital, Harvard Medical School, Harvard Institute of Medicine, Boston, Massachusetts, United States of America.
authors.forename: Jakub
authors.initials: J
authors.lastname: Godlewski
authors.affiliation: Department of Cell Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.
authors.forename: Mirosław
authors.initials: M
authors.lastname: Zarębski
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, Wrocław University of Technology, Wrocław, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Ożyhar
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002522:Chlorocebus aethiops; D029721:Drosophila Proteins / Q000737:chemistry* / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000187:drug effects / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D007605:Juvenile Hormones / Q000494:pharmacology; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects; D020539:Sequence Analysis, Protein; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D029721:Drosophila Proteins; D007605:Juvenile Hormones; D050505:Mutant Proteins; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals; D014157:Transcription Factors; C543965:gce protein, Drosophila