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Título: | Cloning, expression and characterization of mouse spermatid specific thioredoxin-1 gene and protein |
Autor: | Jiménez, Alberto CSIC ORCID CVN; Oko, Richard; Gustafsson, Jan-Åke; Spyrou, Giannis; Pelto-Huikko, Markku; Miranda-Vizuete, Antonio CSIC ORCID | Palabras clave: | Fibrous sheath Sperm Testis Thioredoxin |
Fecha de publicación: | ago-2002 | Editor: | Oxford University Press | Citación: | Human Molecular Genetics 8(8): 710-718 (2002) | Resumen: | Thioredoxins are proteins that participate in different cellular processes via redox-mediated reactions. For humans, we have recently described two novel members of this family that display a male germ cell specific expression pattern, named spermatid specific thioredoxin (Sptrx-1 and Sptrx-2 respectively). We report here the cloning and characterization of the mouse Sptrx-1 gene and protein, which are similar to those described for the human orthologue. The mouse Sptrx-1 open reading frame encodes for a protein of 462 aa composed of an N-terminal repetitive domain of a 15 residue motif followed by a C-terminal domain typical of thioredoxins. The mouse Sptrx-1 gene sequence is interrupted by only one intron of 525 bp located in the 5′-UTR, and using fluorescence in-situ hybridization we have mapped its chromosomal location to 17E1.2–1.3. Northern blot analysis identified the testis as the only tissue expressing mouse Sptrx-1 mRNA, and by in-situ hybridization we found a strong labelling in the testicular seminiferous tubules, mostly in the round spermatids. Affinity purified antibodies against human Sptrx-1 crossreacted well with the mouse protein confirming its expression in seminiferous tubules at the later stages of spermiogenesis. Recombinant mouse Sptrx-1 displayed protein disulphide reducing activity in an enzymatic assay coupled to NADPH and thioredoxin reductase. The availability of the mouse Sptrx-1 gene sequence is the first step towards the generation of knock-out mice, whose characterization will provide significant information regarding the in-vivo function of Sptrx-1 and its possible implication in several sperm anomalies. | Descripción: | 31 páginas, 5 figuras. | Versión del editor: | http://dx.doi.org/10.1093/molehr/8.8.710 | URI: | http://hdl.handle.net/10261/47336 | DOI: | 10.1093/molehr/8.8.710 | ISSN: | 1360-9947 | E-ISSN: | 1460-2407 |
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