Evgeniya N Andreyeva1, Anna A Ogienko2,3, Tatiana D Dubatolova2, Anastasiya L Oshchepkova2,4, Elena N Kozhevnikova2, Anton V Ivankin2, Gera A Pavlova2, Sergei A Kopyl2, Alexey V Pindyurin5,6. 1. Institute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia. andreeva@mcb.nsc.ru. 2. Institute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia. 3. Novosibirsk State University, Novosibirsk, 630090, Russia. 4. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia. 5. Institute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia. a.pindyurin@mcb.nsc.ru. 6. Novosibirsk State University, Novosibirsk, 630090, Russia. a.pindyurin@mcb.nsc.ru.
Abstract
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.
BACKGROUND: Expression of the CNDP2 gene is frequently up- or down-regulated in different types of humancancers. 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.
Authors: Christine Karlsson; Ahmed M Korayem; Christoph Scherfer; Olga Loseva; Mitchell S Dushay; Ulrich Theopold Journal: J Biol Chem Date: 2004-10-04 Impact factor: 5.157
Authors: Anusri Tripathi; Chialin King; Antonio de la Morenas; Victoria Kristina Perry; Bohdana Burke; Gregory A Antoine; Erwin F Hirsch; Maureen Kavanah; Jane Mendez; Michael Stone; Norman P Gerry; Marc E Lenburg; Carol L Rosenberg Journal: Int J Cancer Date: 2008-04-01 Impact factor: 7.396
Authors: Evgeniya N Andreyeva; Travis J Bernardo; Tatyana D Kolesnikova; Xingwu Lu; Lyubov A Yarinich; Boris A Bartholdy; Xiaohan Guo; Olga V Posukh; Sean Healton; Michael A Willcockson; Alexey V Pindyurin; Igor F Zhimulev; Arthur I Skoultchi; Dmitry V Fyodorov Journal: Genes Dev Date: 2017-03-15 Impact factor: 11.361
Authors: Anton Strunov; Lidiya V Boldyreva; Evgeniya N Andreyeva; Gera A Pavlova; Julia V Popova; Alena V Razuvaeva; Alina F Anders; Fioranna Renda; Alexey V Pindyurin; Maurizio Gatti; Elena Kiseleva Journal: BMC Biol Date: 2018-06-15 Impact factor: 7.431