Dual role of histone variant H3.3B in spermatogenesis: positive regulation of piRNA transcription and implication in X-chromosome inactivation

Emeline Fontaine; Christophe Papin; Guillaume Martinez; Stéphanie Le Gras; Roland Abi Nahed; Patrick Héry; Thierry Buchou; Khalid Ouararhni; Bertrand Favier; Thierry Gautier; Jamal S.M. Sabir; Matthieu Gerard; Jan Bednar; Christophe Arnoult; Stefan Dimitrov; Ali Hamiche

doi:10.1093/nar/gkac541

Dual role of histone variant H3.3B in spermatogenesis: positive regulation of piRNA transcription and implication in X-chromosome inactivation

Emeline Fontaine, Christophe Papin, Guillaume Martinez, Stéphanie Le Gras, Roland Abi Nahed, Patrick Héry, Thierry Buchou, Khalid Ouararhni, Bertrand Favier, Thierry Gautier, Jamal S.M. Sabir, Matthieu Gerard, Jan Bednar, Christophe Arnoult^*, Stefan Dimitrov^*, Ali Hamiche^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

The histone variant H3.3 is encoded by two distinct genes, H3f3a and H3f3b, exhibiting identical amino-acid sequence. H3.3 is required for spermatogenesis, but the molecular mechanism of its spermatogenic function remains obscure. Here, we have studied the role of each one of H3.3A and H3.3B proteins in spermatogenesis. We have generated transgenic conditional knock-out/knock-in (cKO/KI) epitope-tagged FLAG-FLAG-HA-H3.3B (H3.3B^HA) and FLAG-FLAG-HA-H3.3A (H3.3A^HA) mouse lines. We show that H3.3B, but not H3.3A, is required for spermatogenesis and male fertility. Analysis of the molecular mechanism unveils that the absence of H3.3B led to alterations in the meiotic/post-meiotic transition. Genome-wide RNA-seq reveals that the depletion of H3.3B in meiotic cells is associated with increased expression of the whole sex X and Y chromosomes as well as of both RLTR10B and RLTR10B2 retrotransposons. In contrast, the absence of H3.3B resulted in down-regulation of the expression of piRNA clusters. ChIP-seq experiments uncover that RLTR10B and RLTR10B2 retrotransposons, the whole sex chromosomes and the piRNA clusters are markedly enriched of H3.3. Taken together, our data dissect the molecular mechanism of H3.3B functions during spermatogenesis and demonstrate that H3.3B, depending on its chromatin localization, is involved in either up-regulation or down-regulation of expression of defined large chromatin regions.

Original language	English
Pages (from-to)	7350-7366
Number of pages	17
Journal	Nucleic Acids Research
Volume	50
Issue number	13
DOIs	https://doi.org/10.1093/nar/gkac541
Publication status	Published - 22 Jul 2022
Externally published	Yes

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10.1093/nar/gkac541

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Fontaine, E., Papin, C., Martinez, G., Le Gras, S., Nahed, R. A., Héry, P., Buchou, T., Ouararhni, K., Favier, B., Gautier, T., Sabir, J. S. M., Gerard, M., Bednar, J., Arnoult, C., Dimitrov, S., & Hamiche, A. (2022). Dual role of histone variant H3.3B in spermatogenesis: positive regulation of piRNA transcription and implication in X-chromosome inactivation. Nucleic Acids Research, 50(13), 7350-7366. https://doi.org/10.1093/nar/gkac541

@article{882161809997499b99fdb7e9e2e5c600,

title = "Dual role of histone variant H3.3B in spermatogenesis: positive regulation of piRNA transcription and implication in X-chromosome inactivation",

abstract = "The histone variant H3.3 is encoded by two distinct genes, H3f3a and H3f3b, exhibiting identical amino-acid sequence. H3.3 is required for spermatogenesis, but the molecular mechanism of its spermatogenic function remains obscure. Here, we have studied the role of each one of H3.3A and H3.3B proteins in spermatogenesis. We have generated transgenic conditional knock-out/knock-in (cKO/KI) epitope-tagged FLAG-FLAG-HA-H3.3B (H3.3BHA) and FLAG-FLAG-HA-H3.3A (H3.3AHA) mouse lines. We show that H3.3B, but not H3.3A, is required for spermatogenesis and male fertility. Analysis of the molecular mechanism unveils that the absence of H3.3B led to alterations in the meiotic/post-meiotic transition. Genome-wide RNA-seq reveals that the depletion of H3.3B in meiotic cells is associated with increased expression of the whole sex X and Y chromosomes as well as of both RLTR10B and RLTR10B2 retrotransposons. In contrast, the absence of H3.3B resulted in down-regulation of the expression of piRNA clusters. ChIP-seq experiments uncover that RLTR10B and RLTR10B2 retrotransposons, the whole sex chromosomes and the piRNA clusters are markedly enriched of H3.3. Taken together, our data dissect the molecular mechanism of H3.3B functions during spermatogenesis and demonstrate that H3.3B, depending on its chromatin localization, is involved in either up-regulation or down-regulation of expression of defined large chromatin regions.",

author = "Emeline Fontaine and Christophe Papin and Guillaume Martinez and \{Le Gras\}, St{\'e}phanie and Nahed, \{Roland Abi\} and Patrick H{\'e}ry and Thierry Buchou and Khalid Ouararhni and Bertrand Favier and Thierry Gautier and Sabir, \{Jamal S.M.\} and Matthieu Gerard and Jan Bednar and Christophe Arnoult and Stefan Dimitrov and Ali Hamiche",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.",

year = "2022",

month = jul,

day = "22",

doi = "10.1093/nar/gkac541",

language = "English",

volume = "50",

pages = "7350--7366",

journal = "Nucleic Acids Research",

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Fontaine, E, Papin, C, Martinez, G, Le Gras, S, Nahed, RA, Héry, P, Buchou, T, Ouararhni, K, Favier, B, Gautier, T, Sabir, JSM, Gerard, M, Bednar, J, Arnoult, C, Dimitrov, S & Hamiche, A 2022, 'Dual role of histone variant H3.3B in spermatogenesis: positive regulation of piRNA transcription and implication in X-chromosome inactivation', Nucleic Acids Research, vol. 50, no. 13, pp. 7350-7366. https://doi.org/10.1093/nar/gkac541

TY - JOUR

T1 - Dual role of histone variant H3.3B in spermatogenesis

T2 - positive regulation of piRNA transcription and implication in X-chromosome inactivation

AU - Fontaine, Emeline

AU - Papin, Christophe

AU - Martinez, Guillaume

AU - Le Gras, Stéphanie

AU - Nahed, Roland Abi

AU - Héry, Patrick

AU - Buchou, Thierry

AU - Ouararhni, Khalid

AU - Favier, Bertrand

AU - Gautier, Thierry

AU - Sabir, Jamal S.M.

AU - Gerard, Matthieu

AU - Bednar, Jan

AU - Arnoult, Christophe

AU - Dimitrov, Stefan

AU - Hamiche, Ali

PY - 2022/7/22

Y1 - 2022/7/22

N2 - The histone variant H3.3 is encoded by two distinct genes, H3f3a and H3f3b, exhibiting identical amino-acid sequence. H3.3 is required for spermatogenesis, but the molecular mechanism of its spermatogenic function remains obscure. Here, we have studied the role of each one of H3.3A and H3.3B proteins in spermatogenesis. We have generated transgenic conditional knock-out/knock-in (cKO/KI) epitope-tagged FLAG-FLAG-HA-H3.3B (H3.3BHA) and FLAG-FLAG-HA-H3.3A (H3.3AHA) mouse lines. We show that H3.3B, but not H3.3A, is required for spermatogenesis and male fertility. Analysis of the molecular mechanism unveils that the absence of H3.3B led to alterations in the meiotic/post-meiotic transition. Genome-wide RNA-seq reveals that the depletion of H3.3B in meiotic cells is associated with increased expression of the whole sex X and Y chromosomes as well as of both RLTR10B and RLTR10B2 retrotransposons. In contrast, the absence of H3.3B resulted in down-regulation of the expression of piRNA clusters. ChIP-seq experiments uncover that RLTR10B and RLTR10B2 retrotransposons, the whole sex chromosomes and the piRNA clusters are markedly enriched of H3.3. Taken together, our data dissect the molecular mechanism of H3.3B functions during spermatogenesis and demonstrate that H3.3B, depending on its chromatin localization, is involved in either up-regulation or down-regulation of expression of defined large chromatin regions.

AB - The histone variant H3.3 is encoded by two distinct genes, H3f3a and H3f3b, exhibiting identical amino-acid sequence. H3.3 is required for spermatogenesis, but the molecular mechanism of its spermatogenic function remains obscure. Here, we have studied the role of each one of H3.3A and H3.3B proteins in spermatogenesis. We have generated transgenic conditional knock-out/knock-in (cKO/KI) epitope-tagged FLAG-FLAG-HA-H3.3B (H3.3BHA) and FLAG-FLAG-HA-H3.3A (H3.3AHA) mouse lines. We show that H3.3B, but not H3.3A, is required for spermatogenesis and male fertility. Analysis of the molecular mechanism unveils that the absence of H3.3B led to alterations in the meiotic/post-meiotic transition. Genome-wide RNA-seq reveals that the depletion of H3.3B in meiotic cells is associated with increased expression of the whole sex X and Y chromosomes as well as of both RLTR10B and RLTR10B2 retrotransposons. In contrast, the absence of H3.3B resulted in down-regulation of the expression of piRNA clusters. ChIP-seq experiments uncover that RLTR10B and RLTR10B2 retrotransposons, the whole sex chromosomes and the piRNA clusters are markedly enriched of H3.3. Taken together, our data dissect the molecular mechanism of H3.3B functions during spermatogenesis and demonstrate that H3.3B, depending on its chromatin localization, is involved in either up-regulation or down-regulation of expression of defined large chromatin regions.

UR - https://www.scopus.com/pages/publications/85134788123

U2 - 10.1093/nar/gkac541

DO - 10.1093/nar/gkac541

M3 - Article

C2 - 35766398

AN - SCOPUS:85134788123

SN - 0305-1048

VL - 50

SP - 7350

EP - 7366

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 13

ER -

Dual role of histone variant H3.3B in spermatogenesis: positive regulation of piRNA transcription and implication in X-chromosome inactivation

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