title: The Novel Gene CRNDE Encodes a Nuclear Peptide (CRNDEP) Which Is Overexpressed in Highly Proliferating Tissues.
abstract: CRNDE, recently described as the lncRNA-coding gene, is overexpressed at RNA level in human malignancies. Its role in gametogenesis, cellular differentiation and pluripotency has been suggested as well. Herein, we aimed to verify our hypothesis that the CRNDE gene may encode a protein product, CRNDEP. By using bioinformatics methods, we identified the 84-amino acid ORF encoded by one of two CRNDE transcripts, previously described by our research team. This ORF was cloned into two expression vectors, subsequently utilized in localization studies in HeLa cells. We also developed a polyclonal antibody against CRNDEP. Its specificity was confirmed in immunohistochemical, cellular localization, Western blot and immunoprecipitation experiments, as well as by showing a statistically significant decrease of endogenous CRNDEP expression in the cells with transient shRNA-mediated knockdown of CRNDE. Endogenous CRNDEP localizes predominantly to the nucleus and its expression seems to be elevated in highly proliferating tissues, like the parabasal layer of the squamous epithelium, intestinal crypts or spermatocytes. After its artificial overexpression in HeLa cells, in a fusion with either the EGFP or DsRed Monomer fluorescent tag, CRNDEP seems to stimulate the formation of stress granules and localize to them. Although the exact role of CRNDEP is unknown, our preliminary results suggest that it may be involved in the regulation of the cell proliferation. Possibly, CRNDEP also participates in oxygen metabolism, considering our in silico results, and the correlation between its enforced overexpression and the formation of stress granules. This is the first report showing the existence of a peptide encoded by the CRNDE gene.
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Lukasz Michal
authors.initials: LM
authors.lastname: Szafron
authors.affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Balcerak
authors.affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Ewa Anna
authors.initials: EA
authors.lastname: Grzybowska
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Pienkowska-Grela
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Felisiak-Golabek
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Podgorska
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kulesza
authors.affiliation: Neurobiology Center, Laboratory of Imaging Tissue Structure and Function, Nencki Institute of Experimental Biology, Warsaw, Poland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Nowak
authors.affiliation: Multimodal Laboratory of Cell Adhesion and Motility, NanoBioGeo Consortium, Nencki Institute of Experimental Biology, Warsaw, Poland; Department of Biochemistry, Laboratory Of Molecular Basis of Cell Motility, Nencki Institute of Experimental Biology, Warsaw, Poland.
authors.forename: Pawel
authors.initials: P
authors.lastname: Pomorski
authors.affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Juliusz
authors.initials: J
authors.lastname: Wysocki
authors.affiliation: The Institute of Radioelectronics, Warsaw University of Technology, Warsaw, Poland.
authors.forename: Tymon
authors.initials: T
authors.lastname: Rubel
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Dansonka-Mieszkowska
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Bozena
authors.initials: B
authors.lastname: Konopka
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Martyna
authors.initials: M
authors.lastname: Lukasik
authors.affiliation: Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.
authors.forename: Jolanta
authors.initials: J
authors.lastname: Kupryjanczyk
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000235:genetics; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics*; D049109:Cell Proliferation / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010455:Peptides / Q000235:genetics*; D062085:RNA, Long Noncoding / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D013090:Spermatocytes / Q000378:metabolism
chemical_list: D000596:Amino Acids; D010455:Peptides; D062085:RNA, Long Noncoding; D034741:RNA, Small Interfering