Identification of a new mouse sperm acrosome-associated protein

Korfanty, Joanna; Toma, Agnieszka; Wojtas, Aleksandra; Rusin, Aleksandra; Vydra, Natalia; Widłak, Wiesława

doi:10.1530/rep-11-0270

Cited by 14 publications

(11 citation statements)

References 29 publications

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“…Moreover, mass spectrometry detected increased abundance of STUB1 and heat shock protein 2 (HSPA2) in Chr18 ∆/∆ mature sperm. Decreased expression of HSPA2 in meiotic and post-meiotic germ cells has been reported in normal mice [45][46][47] , but we observed HSPA2 retention in Chr18 ∆/∆ mature sperm (Fig. 5e, f).…”

Section: Resultssupporting

confidence: 68%

Sperm acrosome overgrowth and infertility in mice lacking chromosome 18 pachytene piRNA

Choi

Zheng-pin

Dean

2020

Preprint

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AbstractpiRNAs are germline-specific, small non-coding RNAs required to maintain genome integrity and preserve RNA homeostasis during male gametogenesis. In murine adult testes, the highest levels of piRNAs are present in the pachytene stage of meiosis, but their mode of action and function remains incompletely understood. We previously reported that BTBD18 binds to 50 pachytene piRNA-producing loci. Here we show that spermatozoa in gene-edited mice lacking a BTBD18 targeted pachytene piRNA cluster on Chr18 have severe sperm head dysmorphology, poor motility, impaired acrosome exocytosis and are sterile. The absence of Chr18 piRNA results in an imbalance of heat shock proteins associated with renaturing proteins and the ubiquitin-proteasome system involved with protein degradation. The mutant phenotype arises from aberrant formation of proacrosomal vesicles, distortion of the trans-Golgi network, and up-regulation of GOLGA2 associated with acrosome dysgenesis. Collectively, our findings reveal central role of pachytene piRNAs in controlling spermiogenesis and male fertility.

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Section: Resultssupporting

confidence: 68%

Sperm acrosome overgrowth and infertility in mice lacking chromosome 18 pachytene piRNA

Choi

Zheng-pin

Dean

2020

Preprint

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“…Extraction of total RNA, purification from DNA contamination, synthesis of cDNA and RT-PCR were performed as described in [ 20 ]. For RT-PCR 1–2 μl of cDNA template was used and 25–35 cycles were applied depending on the primers set.…”

Section: Methodsmentioning

confidence: 99%

Overexpression of heat shock transcription factor 1 enhances the resistance of melanoma cells to doxorubicin and paclitaxel

et al. 2013

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BackgroundHeat Shock Transcription Factor 1 (HSF1) is activated under stress conditions. In turn, it induces expression of Heat Shock Proteins (HSPs), which are well-known regulators of protein homeostasis. Elevated levels of HSF1 and HSPs were observed in many types of tumors. The aim of the present study was to determine whether HSF1 could have an effect on the survival of cancer cells treated with chemotherapeutic cytotoxic agents.MethodsWe constructed mouse (B16F10) and human (1205Lu, WM793B) melanoma cells overexpressing full or mutant form of human HSF1: a constitutively active one with a deletion in regulatory domain or a dominant negative one with a deletion in the activation domain. The impact of different forms of HSF1 on the expression of HSP and ABC genes was studied by RT-PCR and Western blotting. Cell cultures were treated with increasing amounts of doxorubicin, paclitaxel, cisplatin, vinblastine or bortezomib. Cell viability was determined by MTT, and IC50 was calculated. Cellular accumulation of fluorescent dyes and side population cells were studied using flow cytometry.ResultsCells overexpressing HSF1 and characterized by increased HSPs accumulation were more resistant to doxorubicin or paclitaxel, but not to cisplatin, vinblastine or bortezomib. This resistance correlated with the enhanced efflux of fluorescent dyes and the increased number of side population cells. The expression of constitutively active mutant HSF1, also resulting in HSPs overproduction, did not reduce the sensitivity of melanoma cells to drugs, unlike in the case of dominant negative form expression. Cells overexpressing a full or dominant negative form of HSF1, but not a constitutively active one, had higher transcription levels of ABC genes when compared to control cells.ConclusionsHSF1 overexpression facilitates the survival of melanoma cells treated with doxorubicin or paclitaxel. However, HSF1-mediated chemoresistance is not dependent on HSPs accumulation but on an increased potential for drug efflux by ABC transporters. Direct transcriptional activity of HSF1 is not necessary for increased expression of ABC genes, which is probably mediated by HSF1 regulatory domain.

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“…In addition to well-known proteins involved in fertilization, enzymes (proteases, glycosidases, esterases, acid phosphatases, aryl sulfatases), antigens, and various ZP-binding proteins [5], acrosome contains some less-characterized proteins grouped as sperm acrosome associated (SPACA) family proteins. This family has been created based on the cellular localization of its members, but the functions of some of these proteins are still not known [6]. …”

Section: Introductionmentioning

confidence: 99%

“…A novel protein SPACA7 that localizes to the acrosome was described in 2012 by Korfanty and colleagues [6]. Thus new proteins localizing to acrosome are still being discovered.…”

Section: Introductionmentioning

confidence: 99%

A mouse protein that localizes to acrosome and sperm tail is regulated by Y-chromosome

et al. 2013

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BackgroundAcrosomal proteins play crucial roles in the physiology of fertilization. Identification of proteins localizing to the acrosome is fundamental to the understanding of its contribution to fertilization. Novel proteins are still being reported from acrosome. In order to capture yet unreported proteins localizing to acrosome in particular and sperm in general, 2D-PAGE and mass spectrometry analysis of mouse sperm proteins was done.ResultsOne of the protein spots identified in the above study was reported in the NCBI database as a hypothetical protein from Riken cDNA 1700026L06 that localizes to chromosome number 2. Immunofluorescence studies using the antibody raised in rabbit against the recombinant protein showed that it localized to mouse acrosome and sperm tail. Based on the localization of this protein, it has been named mouse acrosome and sperm tail protein (MAST, [Q7TPM5 (http://www.ncbi.nlm.nih.gov/protein/Q7TPM5)]). This protein shows 96% identity to the rat spermatid specific protein RSB66. Western blotting showed that MAST is expressed testis-specifically. Co-immunoprecipitation studies using the MAST antibody identified two calcium-binding proteins, caldendrin and calreticulin as interacting partners of MAST. Caldendrin and calreticulin genes localize to mouse chromosomes 5 and 8 respectively. In a Yq-deletion mutant mouse, that is subfertile and has a deletion of 2/3rd of the long arm of the Y chromosome, MAST failed to localize to the acrosome. Western blot analysis however, revealed equal expression of MAST in the testes of wild type and mutant mice. The acrosomal calcium-binding proteins present in the MAST IP-complex were upregulated in sperms of Yq-del mice.ConclusionsWe have identified a mouse acrosomal protein, MAST, that is expressed testis specifically. MAST does not contain any known motifs for protein interactions; yet it complexes with calcium-binding proteins localizing to the acrosome. The misexpression of all the proteins identified in a complex in the Yq-del mice invokes the hypothesis of a putative pathway regulated by the Y chromosome. The role of Y chromosome in the regulation of this complex is however not clear from the current study.

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Identification of a new mouse sperm acrosome-associated protein

Cited by 14 publications

References 29 publications

Sperm acrosome overgrowth and infertility in mice lacking chromosome 18 pachytene piRNA

Sperm acrosome overgrowth and infertility in mice lacking chromosome 18 pachytene piRNA

Overexpression of heat shock transcription factor 1 enhances the resistance of melanoma cells to doxorubicin and paclitaxel

A mouse protein that localizes to acrosome and sperm tail is regulated by Y-chromosome

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