Gene cloning, expression and functional characterization of a proliferation-inducing ligand (APRIL) from hedgehog (Erinaceus europaeus)

• Published in: Molecular biology reports Vol. 39; no. 5; pp. 6123 - 6132
• Main Authors: Cui, Xian-wei, Xiao, Wen, Ji, Chen-bo, Tian, Ai-ying, Zhang, Jie, Zhang, Shuang-quan
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.05.2012; Springer Nature B.V
• Subjects: Amino Acids - metabolism; Animal Anatomy; Animal Biochemistry; Animals; APRIL protein; B-Lymphocytes - cytology; Biomedical and Life Sciences; Cell growth; Cell Proliferation; Cell Survival; Cloning; Cloning, Molecular; Confocal microscopy; Cysteine; Erinaceus europaeus; Flow cytometry; furin; Gene expression; Gene Expression Profiling; Gene Expression Regulation; Hedgehogs - genetics; Histology; Humans; Life Sciences; Lymphocytes B; Mammals; Mice; Mice, Inbred ICR; Microscopy, Confocal; Microscopy, Fluorescence; Molecular biology; Molecular Sequence Data; Morphology; Organ Specificity - genetics; Polymerase chain reaction; Protein Binding; Proteinase; Proteins; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Homology, Amino Acid; Spleen; Spleen - cytology; Structural Homology, Protein; Tumor necrosis factor; Tumor Necrosis Factor Ligand Superfamily Member 13 - chemistry; Tumor Necrosis Factor Ligand Superfamily Member 13 - genetics; Tumor Necrosis Factor Ligand Superfamily Member 13 - metabolism; GFP; Homology; hedgehog APRIL; Real-time PCR; B-cell survival
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-011-1428-4

Here we describe the identification of the hedgehog Erinaceus europaeus homologue of a proliferation-inducing ligand (APRIL) of the TNF family (designated heAPRIL). Hedgehog APRIL contains two cysteine residues (Cys 196 and Cys 211 ), a furin protease cleavage site and a conserved putative N -glycosylation site (Asn 124 ). Real-time quantitative PCR (qPCR) analysis revealed that heAPRIL could be detected in various tissues. MTT assays and flow cytometric analysis revealed that Nus-hesAPRIL and hesAPRIL could promote the survival/proliferation of splenic B cells . Laser scanning confocal microscopy analysis showed GFP-hesAPRIL could successfully bind to the APRIL receptors of lymphocytes.