TY - JOUR

T1 - A toolset to study functions of Cytosolic non-specific dipeptidase 2 (CNDP2) using Drosophila as a model organism

AU - Andreyeva, Evgeniya N.

AU - Ogienko, Anna A.

AU - Dubatolova, Tatiana D.

AU - Oshchepkova, Anastasiya L.

AU - Kozhevnikova, Elena N.

AU - Ivankin, Anton V.

AU - Pavlova, Gera A.

AU - Kopyl, Sergei A.

AU - Pindyurin, Alexey V.

N1 - Publisher Copyright: © 2019 The Author(s).

PY - 2019/3/18

Y1 - 2019/3/18

N2 - Background: Expression of the CNDP2 gene is frequently up- or down-regulated in different types of human cancers. However, how the product of this gene is involved in cell growth and proliferation is poorly understood. Moreover, our knowledge of the functions of the CNDP2 orthologs in well-established model organisms is scarce. In particular, the function of the D. melanogaster ortholog of CNDP2, encoded by the CG17337 gene (hereafter referred to as dCNDP2), is still unknown. Results: This study was aimed at developing a set of genetic and molecular tools to study the roles of dCNDP2. We generated a dCNDP2 null mutation (hereafter dCNDP2) using CRISPR/Cas9-mediated homologous recombination (HR) and found that the dCNDP2 mutants are homozygous viable, morphologically normal and fertile. We also generated transgenic fly lines expressing eGFP-tagged and non-tagged dCNDP2 protein, all under the control of the UAS promoter, as well as polyclonal antibodies specific to dCNDP2. Using these tools, we demonstrate that only one of the two predicted dCNDP2 isoforms is expressed throughout the different tissues tested. dCNDP2 was detected in both the cytoplasm and the nucleus, and was found to be associated with multiple sites in the salivary gland polytene chromosomes. Conclusions: The dCNDP2 gene is not essential for fly viability under standard laboratory conditions. The subcellular localization pattern of dCNDP2 suggests that this protein might have roles in both the cytoplasm and the nucleus. The genetic and molecular tools developed in this study will allow further functional characterization of the conserved CNDP2 protein using D. melanogaster as a model system.

AB - Background: Expression of the CNDP2 gene is frequently up- or down-regulated in different types of human cancers. However, how the product of this gene is involved in cell growth and proliferation is poorly understood. Moreover, our knowledge of the functions of the CNDP2 orthologs in well-established model organisms is scarce. In particular, the function of the D. melanogaster ortholog of CNDP2, encoded by the CG17337 gene (hereafter referred to as dCNDP2), is still unknown. Results: This study was aimed at developing a set of genetic and molecular tools to study the roles of dCNDP2. We generated a dCNDP2 null mutation (hereafter dCNDP2) using CRISPR/Cas9-mediated homologous recombination (HR) and found that the dCNDP2 mutants are homozygous viable, morphologically normal and fertile. We also generated transgenic fly lines expressing eGFP-tagged and non-tagged dCNDP2 protein, all under the control of the UAS promoter, as well as polyclonal antibodies specific to dCNDP2. Using these tools, we demonstrate that only one of the two predicted dCNDP2 isoforms is expressed throughout the different tissues tested. dCNDP2 was detected in both the cytoplasm and the nucleus, and was found to be associated with multiple sites in the salivary gland polytene chromosomes. Conclusions: The dCNDP2 gene is not essential for fly viability under standard laboratory conditions. The subcellular localization pattern of dCNDP2 suggests that this protein might have roles in both the cytoplasm and the nucleus. The genetic and molecular tools developed in this study will allow further functional characterization of the conserved CNDP2 protein using D. melanogaster as a model system.

KW - CG17337

KW - CN2

KW - CNDP2

KW - CPGL

KW - CRISPR/Cas9

KW - Drosophila melanogaster

KW - Tumor suppressor gene

KW - IDENTIFICATION

KW - GENOME

KW - CARNOSINASE

KW - CARBOXYPEPTIDASE

KW - SEQUENCE

KW - GROWTH

KW - PROTEOMIC ANALYSIS

KW - PROTEINS

KW - MELANOGASTER

KW - CYCLE

UR - http://www.scopus.com/inward/record.url?scp=85063157868&partnerID=8YFLogxK

U2 - 10.1186/s12863-019-0726-z

DO - 10.1186/s12863-019-0726-z

M3 - Article

C2 - 30885138

AN - SCOPUS:85063157868

VL - 20

SP - 31

JO - BMC Genetics

JF - BMC Genetics

SN - 1471-2156

IS - Suppl 1

M1 - 31

ER -