Cloning and characterization of a novel gene which encodes a protein interacting with the mitosis-associated kinase-like protein NTKL.
NTKL is an evolutionarily conserved kinase-like protein. The cell-cycle-dependent centrosomal localization of NTKL suggested that it was involved in centrosome-related cellular function. The mouse NTKL protein is highly homologous with human NTKL. A novel mouse protein was identified as an NTKL-binding protein (NTKL-BP1) by yeast two-hybrid screening, and the full-length cDNA was amplified based on the result of a sequence data analysis cloning strategy. The full-length cDNA sequence of the NTKL-BP1 gene consists of 2,537 bp, which encode 368 amino acids. A database search revealed that homologues of NTKL-BP1 exist in different organisms, including Arabidopsis thaliana, Drosophila melanogaster, Plasmodium falciparum, Geobacter metallireducens, Anopheles gambiae and human. It suggests that NTKL-BP1 is an evolutionarily conserved protein. The expression of NTKL-BP1 was observed in multiple normal mouse tissues. The interaction of the two proteins was confirmed by co-immunoprecipitation. Moreover, immunofluorescent staining indicated that NTKL and NTKL-BP1 were all localized in the cytoplasm.
Di
Yujun
Y
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
Li
Jinjun
J
State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, 200032, Shanghai, PR China.
Fang
Ji
J
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
Xu
Zhigang
Z
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
He
Xianghuo
X
State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, 200032, Shanghai, PR China.
Zhang
Fengrui
F
State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, 200032, Shanghai, PR China.
Ling
Jiqiang
J
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
Li
Xinrui
X
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
Xu
Dongbin
D
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
Li
Li
L
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
Li
Yu-Yang
YY
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China.
Huo
Keke
K
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 200433, Shanghai, PR China. kkhuo@fudan.edu.cn.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics*; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D030541:Databases, Genetic; D019085:Fluorescent Antibody Technique, Direct; D000077614:Golgi Matrix Proteins; D051379:Mice; D008938:Mitosis; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics*; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins
D002352:Carrier Proteins; D000077614:Golgi Matrix Proteins; C000627657:Gorab protein, mouse; D033921:Vesicular Transport Proteins; D011494:Protein Kinases