Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>)
In mammals, the testis-specific serine/threonine kinase (TSSK) is essential for spermatogenesis and male fertility. TSSK4 belongs to the family of the testis-specific serine/threonine-protein kinase (TSSK), with a crucial role in spermatogenesis. This study aimed to analyze the variable spliceosome...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-05-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/12/11/1380 |
| _version_ | 1797494406922633216 |
|---|---|
| author | Xingdong Wang Jie Pei Lin Xiong Shaoke Guo Mengli Cao Yandong Kang Pengjia Bao Xiaoyun Wu Min Chu Chunnian Liang Ping Yan Xian Guo |
| author_facet | Xingdong Wang Jie Pei Lin Xiong Shaoke Guo Mengli Cao Yandong Kang Pengjia Bao Xiaoyun Wu Min Chu Chunnian Liang Ping Yan Xian Guo |
| author_sort | Xingdong Wang |
| collection | DOAJ |
| description | In mammals, the testis-specific serine/threonine kinase (TSSK) is essential for spermatogenesis and male fertility. TSSK4 belongs to the family of the testis-specific serine/threonine-protein kinase (TSSK), with a crucial role in spermatogenesis. This study aimed to analyze the variable spliceosome of the TSSK4 gene in the yak for understanding the regulatory function of the TSSK4 spliceosome in yak testis development using PCR amplification and cloning techniques. The GST pull-down was used for pulling down the protein interacting with TSSK4, and then the protein interacting with TSSK4 was identified using LC–MS/MS. The results of the PCR amplification demonstrated multiple bands of the TSSK4 gene in the yak. The cloning and sequencing yielded a total of six alternative spliceosomes, which included only two alternative spliceosomes before sexual maturity and four alternative spliceosomes after sexual maturity. The sub-cells of the alternative spliceosomes were found to localize in the nucleus before sexual maturity and in the cytoplasm after sexual maturity. The LC–MS/MS analysis of the alternative spliceosome with the highest expression after sexual maturity yielded a total of 223 interacting proteins. The enrichment analysis of the 223 interacting proteins revealed these proteins participate in biological processes, cell composition, and molecular functions. The KEGG analysis indicated that the TSSK4-interacting protein participates in the estrogen signaling pathways, tight junctions, endoplasmic reticulum protein processing, and other signaling pathways. This study cloned the six alternative spliceosomes of the TSSK4 gene laying the foundation for studying the function of each spliceosome in the future. |
| first_indexed | 2024-03-10T01:33:50Z |
| format | Article |
| id | doaj.art-b193db6990074750a3f2d66061b88a3b |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T01:33:50Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-b193db6990074750a3f2d66061b88a3b2023-11-23T13:37:22ZengMDPI AGAnimals2076-26152022-05-011211138010.3390/ani12111380Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>)Xingdong Wang0Jie Pei1Lin Xiong2Shaoke Guo3Mengli Cao4Yandong Kang5Pengjia Bao6Xiaoyun Wu7Min Chu8Chunnian Liang9Ping Yan10Xian Guo11Key Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaKey Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, ChinaIn mammals, the testis-specific serine/threonine kinase (TSSK) is essential for spermatogenesis and male fertility. TSSK4 belongs to the family of the testis-specific serine/threonine-protein kinase (TSSK), with a crucial role in spermatogenesis. This study aimed to analyze the variable spliceosome of the TSSK4 gene in the yak for understanding the regulatory function of the TSSK4 spliceosome in yak testis development using PCR amplification and cloning techniques. The GST pull-down was used for pulling down the protein interacting with TSSK4, and then the protein interacting with TSSK4 was identified using LC–MS/MS. The results of the PCR amplification demonstrated multiple bands of the TSSK4 gene in the yak. The cloning and sequencing yielded a total of six alternative spliceosomes, which included only two alternative spliceosomes before sexual maturity and four alternative spliceosomes after sexual maturity. The sub-cells of the alternative spliceosomes were found to localize in the nucleus before sexual maturity and in the cytoplasm after sexual maturity. The LC–MS/MS analysis of the alternative spliceosome with the highest expression after sexual maturity yielded a total of 223 interacting proteins. The enrichment analysis of the 223 interacting proteins revealed these proteins participate in biological processes, cell composition, and molecular functions. The KEGG analysis indicated that the TSSK4-interacting protein participates in the estrogen signaling pathways, tight junctions, endoplasmic reticulum protein processing, and other signaling pathways. This study cloned the six alternative spliceosomes of the TSSK4 gene laying the foundation for studying the function of each spliceosome in the future.https://www.mdpi.com/2076-2615/12/11/1380yakTSSK4alternative spliceosomesGST pull-downLC–MS/MStestis |
| spellingShingle | Xingdong Wang Jie Pei Lin Xiong Shaoke Guo Mengli Cao Yandong Kang Pengjia Bao Xiaoyun Wu Min Chu Chunnian Liang Ping Yan Xian Guo Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>) Animals yak TSSK4 alternative spliceosomes GST pull-down LC–MS/MS testis |
| title | Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>) |
| title_full | Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>) |
| title_fullStr | Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>) |
| title_full_unstemmed | Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>) |
| title_short | Identification of the TSSK4 Alternative Spliceosomes and Analysis of the Function of the TSSK4 Protein in Yak (<i>Bos grunniens</i>) |
| title_sort | identification of the tssk4 alternative spliceosomes and analysis of the function of the tssk4 protein in yak i bos grunniens i |
| topic | yak TSSK4 alternative spliceosomes GST pull-down LC–MS/MS testis |
| url | https://www.mdpi.com/2076-2615/12/11/1380 |
| work_keys_str_mv | AT xingdongwang identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT jiepei identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT linxiong identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT shaokeguo identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT menglicao identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT yandongkang identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT pengjiabao identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT xiaoyunwu identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT minchu identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT chunnianliang identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT pingyan identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi AT xianguo identificationofthetssk4alternativespliceosomesandanalysisofthefunctionofthetssk4proteininyakibosgrunniensi |