Ran binding protein RanBP1 in zebrafish embryonic development

• Published in: Molecular reproduction and development Vol. 59; no. 3; pp. 235 - 248
• Main Authors: Mangos, Steve, Vanderbeld, Barbara, Krawetz, Roman, Sudol, Krzysztof, Kelly, Gregory M.
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 01.07.2001; Wiley-Liss
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Biotechnology; Cloning, Molecular; Danio rerio; embryogenesis; Freshwater; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Humans; In Situ Hybridization; Methods. Procedures. Technologies; Microinjections; Molecular cloning; Molecular Sequence Data; Nuclear Proteins - chemistry; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Phenotype; Precipitin Tests; Ran; ran GTP-Binding Protein - chemistry; ran GTP-Binding Protein - genetics; ran GTP-Binding Protein - metabolism; RanBP1; RanBP1 gene; Reverse Transcriptase Polymerase Chain Reaction; RNA interference; RNA, Messenger - metabolism; Xenopus laevis - metabolism; zebrafish; Zebrafish - embryology; Embryonic development; Molecular hybridization; RNA; Enzyme; Triphosphoric monoester hydrolases; Xenopus laevis; Amphibia; dGTPase; Esterases; Salientia; Gene expression; Vertebrata; Hydrolases; Molecular cloning; Aminoacid sequence
• ISSN: 1040-452X; 1098-2795
• DOI: 10.1002/mrd.1028

Ran, which functions in nucleocytoplasmic transport and mitosis, binds to and is regulated in part by RanBP1. We have identified a zebrafish RanBP1 cDNA and report that it encodes for a polypeptide of 233 amino acids with considerable similarity to human and Xenopus RanBP1, despite the fact that it is 10% longer due to an extension at its carboxy terminus. RanBP1 mRNA is present as a maternal transcript and is expressed ubiquitously throughout the developing embryo. At the protein level, RanBP1 is present at all embryonic stages. Surprisingly, the ectopic overexpression of the protein had no obvious effect on embryogenesis. Attempts were also made to down‐regulate RanBP1 activity by RNA interference. Injecting double‐stranded RNA augmented both the mortality rate and the frequency of induced defects. Specific defects accompanied by changes in RanBP1 expression were not seen, leading us to propose that RNAi is not a reliable method for deregulating the activity of constitutively expressed genes, like RanBP1, in zebrafish. Mol. Reprod. Dev. 59:235–248, 2001. © 2001 Wiley‐Liss, Inc.