A global screen for testicular proteins and RNAs interacting with STRBP

Búsqueda global de proteínas y ARN testiculares que interactúan con STRBP

Background: Spermatid perinuclear RNA-binding protein (STRBP) facilitates RNAs transportation and translational activation during spermatid development. STRBP deficiency impairs male fertility and causes premature death of mouse offspring. However, the mechanisms underlying the regulation of spermiogenesis remain elusive. Methods: RNA immunoprecipitation sequencing and immunoprecipitation, followed by liquid chromatography-mass spectrometry, were used to explore how RNAs and proteins interact with STRBP. Results: These results suggest that STRBP is involved in spermiogenesis, mainly through precursor messenger RNA (mRNA) maturation by the spliceosome, RNAs transportation, translation-activating or translation-inactivating processes, and interaction with long noncoding RNAs. The transport action exerted by STRBP involves interactions with RNAs and proteins, including centromere protein E (Cenpe), dynein axonemal intermediate chain 3 (Dnai3), kinesin family member 23 (Kif23), kinesin family member 20b (Kif20b), dynein axonemal intermediate chain 1 (Dnai1), tubulin tyrosine ligase like 3 (Ttll3), DExH-Box helicase 9 (DHX9) and heterogeneous nuclear ribonucleoprotein A2/B1 (HNRNPA2B1). Transformer 2 beta homolog (TRA2B), HNRNPA2B1, serine and arginine rich splicing factor 2 (SRSF2), apoptotic chromatin condensation inducer 1 (ACIN1), sin3A associated protein 18 (SAP18), la ribonucleoprotein 7 (Larp7) and transcription elongation regulator 1 (Tcerg1), may mediate nuclear precursor mRNA maturation. Poly(RC) binding protein 2 (PCBP2), DHX9, synaptotagmin binding cytoplasmic RNA interacting protein (SYNCRIP), heterogeneous nuclear ribonucleoprotein D (HNRNPD) and N-acetyltransferase 10 (NAT10) are involved in the translation process, whereas UPF1 RNA helicase and ATPase (UPF1) and ribosomal protein L13a (RPL13A) probably impose a delayed action on mRNA translation. Conclusions: Our study not only provides preliminary evidence on STRBP-regulated processes involved in sperm physiology but also indicates potential candidates for investigating their roles in spermatogenesis.

Resumen

Antecedentes: La proteína de unión al ARN perinuclear de los espermatozoides (STRBP) facilita el transporte de ARN y la activación traslacional durante el desarrollo de los espermatozoides. La deficiencia de STRBP afecta a la fertilidad masculina y causa la muerte prematura de las crías de ratón. Sin embargo, los mecanismos subyacentes a la regulación de la espermiogénesis siguen siendo esquivos. Métodos: En este trabajo se utilizó la secuenciación por inmunoprecipitación de ARN y la inmunoprecipitación, seguidas de cromatografía líquida-espectrometría de masas, para explorar cómo interactúan los ARN y las proteínas con la STRBP. Resultados: Estos resultados sugieren que la STRBP interviene en la espermiogénesis, principalmente a través de la maduración del ARNm precursor por el espliceosoma, el transporte de ARNs, los procesos de activación o desactivación de la traducción y la interacción con ARN no codificantes largos. La acción de transporte ejercida por la STRBP implica interacciones con ARN y proteínas, entre ellas la proteína E del centrómero (Cenpe), la cadena intermedia axonemal 3 de la dineína (Dnai3), el miembro 23 de la familia de la kinesina (Kif23), el miembro 20b de la familia de la kinesina (Kif20b), la cadena intermedia axonemal 1 de la dineína (Dnai1), la tubulina tirosina ligasa como 3 (Ttll3), la helicasa 9 DExH-Box (DHX9) y la ribonucleoproteína nuclear heterogénea A2/B1 (HNRNPA2B1). El homólogo beta del transformador 2 (TRA2B), la HNRNPA2B1, el factor de empalme 2 rico en serina y arginina (SRSF2), el inductor 1 de la condensación apoptótica de la cromatina (ACIN1), la proteína 18 asociada a sin3A (SAP18), la ribonucleoproteína 7 (Larp7) y el regulador 1 de la elongación de la transcripción (Tcerg1), pueden mediar en la maduración del ARNm precursor nuclear. La proteína de unión a poli(RC) 2 (PCBP2), DHX9, la proteína de interacción con el ARN citoplasmático de unión a sinaptotagmina (SYNCRIP), la ribonucleoproteína nuclear heterogénea D (HNRNPD) y la N-acetiltransferasa 10 (NAT10) participan en el proceso de traducción, mientras que la ARN helicasa y ATPasa UPF1 (UPF1) y la proteína ribosómica L13a RPL13A probablemente imponen una acción retardada en la traducción del ARNm. Conclusiones: En conclusión, nuestro estudio no sólo aporta pruebas preliminares sobre los procesos regulados por STRBP implicados en la fisiología espermática, sino que también indica posibles candidatos para investigar sus funciones en la espermatogénesis.

STRBP; Spermiogenesis; RNA transportation; Cytoplasmic translation; lncRNAs; Spermatogenesis

Palabras Clave

STRBP; Espermiogénesis; Transporte de ARN; Traducción citoplasmática; lncARNs; Espermatogénesis

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