Abstract
Spermatogenesis associated 4 (Spata4), a testis-specific and CpG island associated gene, is involved in regulating cell proliferation, differentiation and apoptosis. To obtain insight into the role of Spata4 in cell cycling control, we characterized the promoter region of Spata4 and investigated its transcriptional regulation mechanism. The Spata4 promoter is unidirectional transcribed and possesses multiple transcription start sites. Moreover, we present evidence that regulatory factor X1 (RFX1) could bind the typical 14-bp cis-elements of Spata4 promoter, modulate transcriptional activity and endogenous expression of Spata4, and further regulate the proliferation of Sertoli cells. Overexpression of RFX1 was shown to down-regulate both the promoter activity and mRNA expression of Spata4, whereas knockdown of RFX1 demonstrated the opposite effects. Our studies provide insight into Spata4 gene regulation and imply the potential role of RFX1 in growth of Sertoli cells. RFX1 may have negative effect on cell proliferation of Sertoli cells via modulating Spata4 expression levels by binding the conserved 14-bp cis-elements of Spata4 promoter.
Highlights
Spermatogenesis associated 4 (Spata4) was initially identified in human testes and plays an important role in cryptorchidism development [1]
The mouse Spata4 promoter contains a CpG island approximately 400-bp-long (Figure 1B) with GC % = 54% and ObsCpG/ExpCpG = 0.604, which comply with the accepted definition of CpG island [14]
Transcription of Spata4 initiates with a purine at +1 at a frequency of 98% and with a pyrimidine-purine dinucleotide at position 21, +1 at a frequency of 88%, which is consistent with previous studies that RNA polymerase II-mediated transcription initiation in mammals is preferentially initiated at a pyrimidine-purine dinucleotide at position 21, +1 [31]
Summary
Spermatogenesis associated 4 (Spata4) was initially identified in human testes and plays an important role in cryptorchidism development [1]. Recent studies indicate that Spata modulates cell growth, proliferation and differentiation in various cell types [1,7,8]. Sertoli cells produce essential factors for germ cell development and modulate male fertility [9]. These factors are of great importance in the process of spermatogenesis [10,11]. Sertoli cell growth and proliferation may contribute to maintaining the normal physiological functions of male reproductive system
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