Molecular cloning and characterization of a novel transcript variant of Mtsarg1 gene.
Mtsarg1 (Mus musculus testis and spermatogenesis cell apoptosis-related gene 1) gene with 1103 bp in full length had been cloned previously (GenBank accession number: AF399971, 2002; re-designated as Spata3, Mus musculus spermatogenesis-associated 3, 2007). In the present study, we identified a novel transcript variant of Mtsarg1, named Mtsarg1-beta which is 887 bp in length (GenBank accession numbers: EU259321 and EF546784, 2007, designated as Spata3 variant 4) by reverse transcription-polymerase chain reaction (RT-PCR), cloning and sequencing. Mtsarg1-beta which has high similarity with Mtsarg1 contains an entire open reading frame of 417 bp encoding a protein consisting of 138 amino acids. Mtsarg1-beta protein is a non-secretory protein with a theoretic molecular mass around 14.79 kD and an isoelectric point of 9.74, which shares the 100 N-terminal amino acids with Mtsarg1 followed by 38 amino acids differing from Mtsarg1. Multi-tissue RT-PCR results and Northern blot analysis for adult DBA/2 mice showed that Mtsarg1-beta and Mtsarg1 mRNAs were specifically expressed in testis at high level. RT-PCR results also showed that Mtsarg1-beta and Mtsarg1 mRNAs were not expressed in mouse GC-1 spermatogonia. In situ hybridization revealed that both Mtsarg1 and probably Mtsarg1-beta mRNAs were mainly expressed in mouse spermatocytes. Subcellular localization analysis suggested that Mtsarg1 protein was mainly localized in nucleus while Mtsarg1-beta protein was mainly localized in cytoplasm. All these results indicate that Mtsarg1 and Mtsarg1-beta may play an important role in mouse testicular function and in spermatocyte development.
Li
Li
L
Institute of Human Reproductive and Stem Cell Engineering, Xiangya School of Medicine, Central South University, Changsha, Hunan, People's Republic of China.
Liu
Gang
G
Fu
Jun-Jiang
JJ
Li
Lu-Yun
LY
Tan
Xiao-Jun
XJ
Yang
Sheng
S
Lu
Guang-Xiu
GX
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D019295:Computational Biology; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D017403:In Situ Hybridization; D007438:Introns / Q000235:genetics; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013093:Spermatogonia / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D018076:DNA, Complementary; D020033:Protein Isoforms; D011506:Proteins; D012333:RNA, Messenger; C542262:Tsarg1 protein, mouse