A single amino acid mutation in the mouse MEIG1 protein disrupts a cargo transport system necessary for sperm formation

Wei, Li; Huang, Qian; Zhang, Ling; Liu, Hong; Zhang, David; Yuan, Shuo; Yap, Yitian; Qü, Wei; Shiang, Rita; Song, Shizheng; Hess, Rex A.; Zhang, Zhibing

doi:10.1016/j.jbc.2021.101312

Cited by 7 publications

(19 citation statements)

References 49 publications

(76 reference statements)

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“…Our previous studies demonstrated that MEIG1 and PACRG form a complex in the manchette for protein cargo transport to build the sperm flagellum. We further discovered that four amino acids on the same surface of MEIG1 protein are involved in its interaction with PACRG (28, 29). However, it was not clear how the MEIG1/PACRG complex associates with the manchette microtubules.…”

Section: Discussionmentioning

confidence: 97%

“…MEIG1 was present in the cell bodies of spermatocytes and round spermatids of control mice, and protein localization was not changed in the two cell types in Dnali1 cKO mice (27,29, Figure 9B). Similarly, SPAG16L was present in the cytoplasm of spermatocytes and round spermatids in both control and Dnali1 cKO mice (27,29, Figure 9C). In contrast, while MEIG1 and SPAG16L were present in elongating spermatids of Dnali1 cKO mice, they did not co-localized in the manchette as observed for control mice (Figure 9D, Supplemental Figure 6).…”

Section: Inactivation Of Dnali1 In Male Germ Cells Did Not Change Pro...mentioning

confidence: 95%

“…Localization of MEIG1 and SPAG16L was further examined. MEIG1 was present in the cell bodies of spermatocytes and round spermatids of control mice, and protein localization was not changed in the two cell types in Dnali1 cKO mice (27,29, Figure 9B). Similarly, SPAG16L was present in the cytoplasm of spermatocytes and round spermatids in both control and Dnali1 cKO mice (27,29, Figure 9C).…”

Section: Inactivation Of Dnali1 In Male Germ Cells Did Not Change Pro...mentioning

confidence: 95%

“…We previously discovered that mouse meiosis-expressed gene 1 (MEIG1) and Parkin co-regulated gene (PACRG) form a protein complex in the manchette, a transient microtubule structure localized in the head compartment of sperm cells during differentiation. It was demonstrated that this protein complex controls sperm flagellum formation, with PACRG being an upstream molecule of MEIG1 (26)(27)(28)(29). The MEIG1/PACRG complex was shown to function as part of the cargo transport system, called intra-manchette transport system (IMT) (30), which transports sperm flagella proteins, including SPAG16, for sperm tail assembly.…”

Section: Introductionmentioning

confidence: 99%

“…The Parcg gene is a reverse strand gene located upstream of the Parkin gene, involved in Parkinson's disease (34). Genetic disruption of Pacrg in mouse phenocopies the infertility phenotype of Meig1 knockout mice (27,29,(35)(36)(37)(38). Our previous studies demonstrated that MEIG1 and PACRG form a complex in the manchette for protein cargo transport to build the sperm flagellum.…”

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confidence: 99%

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MEIG1/PACRG associated and non-associated functions of axonemal dynein light intermediate polypeptide 1 (DNALI1) in mammalian spermatogenesis

Yap

Huang

et al. 2022

Preprint

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Axonemal dynein light intermediate polypeptide 1 (DNALI1) was originally cloned from Chlamydomonas reinhardtii in an effort to find motor proteins essential for flagellar motility. Here we report that DNALI1 is a binding partner of parkin co-regulated gene 1 (PACRG), which forms a complex with meiosis expressed gene 1 (MEIG1) in the manchette, a transient and unique structure only present in the elongating spermatids and required for normal spermiogenesis of the male germ cell differentiation process. DNALI1 recruits the PACRG protein in transfected CHO cells, and also stabilizes PACRG in bacteria and transfected mammalian cells. The untagged DNALI1 could also be co-purified with His-tagged PACRG in the gel filtration assay. Immunofluorescence staining on isolated male germ cells revealed that DNALI1 was present in the manchette of elongating spermatids, and colocalized with PACRG in this structure. In Pacrg mutant mice, localization of DNALI1 in the manchette was not changed, suggesting that DNALI1 and PACRG form a complex in the manchette, with DNALI1 being an upstream molecule. Mice deficiency in DNALI1 specifically in male germ cells showed dramatically reduced sperm numbers and were infertile. In addition, majority of the sperm exhibited abnormal morphology including misshapen heads, bent tails and enlarged midpiece, discontinuous accessory structure, and loss of sperm individualization, emphasizing the importance of DNALI1 in sperm development. Examination of testis histology revealed impaired spermiogenesis in the conditional Dnali1 knockout mice. Electron microscopy revealed disrupted ultrastructure in sperm of the Dnali1 mutant mice. Testicular levels of MEIG1, PACRG and SPAG16L proteins were not changed in the Dnali1 mutant mice. However, MEIG1 and SPAG16L were no longer present in the manchette in the absence of DNALI1. These findings demonstrate that DNALI1 is involved in the connection of the MEIG1/PACRG complex to carry cargo proteins along the manchette microtubules for sperm flagella formation. Given that Dnali1 mutant mice showed impaired sperm individualization that was not observed in the MEIG1 nor PACRG-deficient mice, DNALI1 might fulfill other functions beyond its role associated with the MEIG1/PACRG complex. Thus, DNALI1 plays multiple roles in sperm cell differentiation and function.Summary statementAxonemal dynein light intermediate polypeptide 1 (DNALI1) is required for sperm formation and male fertility. It associates with the MEIG1/PACRG complex in the manchette and is involved in a cargo transport system. In addition, it might be related to IFT and sperm individualization.

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Section: Discussionmentioning

confidence: 97%

Section: Inactivation Of Dnali1 In Male Germ Cells Did Not Change Pro...mentioning

confidence: 95%

Section: Inactivation Of Dnali1 In Male Germ Cells Did Not Change Pro...mentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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MEIG1/PACRG associated and non-associated functions of axonemal dynein light intermediate polypeptide 1 (DNALI1) in mammalian spermatogenesis

Yap

Huang

et al. 2022

Preprint

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Molecular dynamics study reveals key disruptors of MEIG1–PACRG interaction

2022

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Interactions between the meiosis‐expressed gene 1 (MEIG1) and Parkin co‐regulated gene (PACRG) protein are critical in the formation of mature sperm cells. Targeting either MEIG1 or PACRG protein could be a contraceptive strategy. The W50A and Y68A mutations on MEIG1 are known to interrupt the MEIG1–PACRG interactions resulting in defective sperm cells. However, the details about how the mutants disrupt the protein–protein binding are not clear. In this study, we reveal insights on MEIG1 and PACRG protein dynamics by applying Gaussian‐accelerated molecular dynamics (GaMD) simulations and post‐GaMD analysis. Our results show that the mutations destabilize the protein–protein interfacial interaction. The effect of the Y68A mutation is more significant than W50A as Y68 forms stronger polar interactions with PACRG. Because both human and mouse models demonstrate similar dynamic properties, the findings from mouse proteins can be applied to the human system. Moreover, we report a potential ligand binding pocket on the MEIG1 and PACRG interaction surface that could be a target for future drug design to inhibit the MEIG1–PACRG interaction. PACRG shows more qualified pockets along the protein–protein interface, implying that it is a better target than MEIG1. Our work provides a fundamental understanding of MEIG1 and PACRG protein dynamics, paving the way for drug discovery in male‐based contraception.

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TCFL5 deficiency impairs the pachytene to diplotene transition during spermatogenesis in the mouse

Galán-Martínez

Berenguer

Maza

et al. 2022

Sci Rep

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Spermatogenesis is a complex, multistep process during which spermatogonia give rise to spermatozoa. Transcription Factor Like 5 (TCFL5) is a transcription factor that has been described expressed during spermatogenesis. In order to decipher the role of TCFL5 during in vivo spermatogenesis, we generated two mouse models. Ubiquitous removal of TCFL5 generated by breeding TCFL5fl/fl with SOX2-Cre mice resulted in sterile males being unable to produce spermatozoa due to a dramatic alteration of the testis architecture presenting meiosis arrest and lack of spermatids. SYCP3, SYCP1 and H1T expression analysis showed that TCFL5 deficiency causes alterations during pachytene/diplotene transition resulting in a meiotic arrest in a diplotene-like stage. Even more, TCFL5 deficient pachytene showed alterations in the number of MLH1 foci and the condensation of the sexual body. In addition, tamoxifen-inducible TCFL5 knockout mice showed, besides meiosis phenotype, alterations in the spermatids elongation process resulting in aberrant spermatids. Furthermore, TCFL5 deficiency increased spermatogonia maintenance genes (Dalz, Sox2, and Dmrt1) but also increased meiosis genes (Syce1, Stag3, and Morc2a) suggesting that the synaptonemal complex forms well, but cannot separate and meiosis does not proceed. TCFL5 is able to bind to the promoter of Syce1, Stag3, Dmrt1, and Syce1 suggesting a direct control of their expression. In conclusion, TCFL5 plays an essential role in spermatogenesis progression being indispensable for meiosis resolution and spermatids maturation.

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A single amino acid mutation in the mouse MEIG1 protein disrupts a cargo transport system necessary for sperm formation

Cited by 7 publications

References 49 publications

MEIG1/PACRG associated and non-associated functions of axonemal dynein light intermediate polypeptide 1 (DNALI1) in mammalian spermatogenesis

MEIG1/PACRG associated and non-associated functions of axonemal dynein light intermediate polypeptide 1 (DNALI1) in mammalian spermatogenesis

Molecular dynamics study reveals key disruptors of MEIG1–PACRG interaction

TCFL5 deficiency impairs the pachytene to diplotene transition during spermatogenesis in the mouse

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