Cytoplasmic pool of U1 spliceosome protein SNRNP70 shapes the axonal transcriptome and regulates motor connectivity

Nikolas Nikolaou; Patricia Gordon; Fursham Hamid; Richard Taylor; Joshua Lloyd-Jones; Eugene Makeyev; Corinne Houart

doi:10.1016/j.cub.2022.10.048

Cytoplasmic pool of U1 spliceosome protein SNRNP70 shapes the axonal transcriptome and regulates motor connectivity

Nikolas Nikolaou, Patricia Gordon, Fursham Hamid, Richard Taylor, Joshua Lloyd-Jones, Eugene Makeyev, Corinne Houart

Department of Life Sciences

King's College London

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

11 Citations (SciVal)

Abstract

Regulation of pre-mRNA splicing and polyadenylation plays a profound role in neurons by diversifying the proteome and modulating gene expression in response to physiological cues. Although most of the pre-mRNA processing is thought to occur in the nucleus, numerous splicing regulators are also found in neurites. Here, we show that U1-70K/SNRNP70, a component of the major spliceosome, localizes in RNA-associated granules in zebrafish axons. We identify the extra-nuclear SNRNP70 as an important regulator of motor axonal growth, nerve-dependent acetylcholine receptor (AChR) clustering, and neuromuscular synaptogenesis. This cytoplasmic pool has a protective role for a limited number of transcripts regulating their abundance and trafficking inside axons. Moreover, non-nuclear SNRNP70 regulates splice variants of transcripts such as agrin, thereby controlling synapse formation. Our results point to an unexpected, yet essential, function of non-nuclear SNRNP70 in axonal development, indicating a role of spliceosome proteins in cytoplasmic RNA metabolism during neuronal connectivity.

Original language	English
Pages (from-to)	5099-5115.e8
Journal	Current Biology
Volume	32
Issue number	23
DOIs	https://doi.org/10.1016/j.cub.2022.10.048
Publication status	Published - 5 Dec 2022

Bibliographical note

Funding Information:
We thank Prof. David Stanek for kindly giving us the human SNRNP70 construct, Prof. Martin Meyer for the HuC:Gal4 construct, Triona Fielding for technical support throughout the work, and Dr. Rachel Moore and Prof. Jon Clarke for assistance in using the Zeiss Airyscan confocal imaging system. We also thank the staff at King’s College London (Guy’s Campus) and the University of Bath fish facilities for their fish husbandry and care. This study was supported by the Biotechnology and Biological Sciences Research Council ( BB/P001599/1 to C.H. and BB/M007103/1 , BB/R001049/1 , and BB/V006258/1 to E.V.M.), the Wellcome Trust (Tech Dev. WT093389 and Investigator Award WT220861/Z to C.H.), and a University of Bath start-up package awarded to N.N.

Funding Information:
We thank Prof. David Stanek for kindly giving us the human SNRNP70 construct, Prof. Martin Meyer for the HuC:Gal4 construct, Triona Fielding for technical support throughout the work, and Dr. Rachel Moore and Prof. Jon Clarke for assistance in using the Zeiss Airyscan confocal imaging system. We also thank the staff at King's College London (Guy's Campus) and the University of Bath fish facilities for their fish husbandry and care. This study was supported by the Biotechnology and Biological Sciences Research Council (BB/P001599/1 to C.H. and BB/M007103/1, BB/R001049/1, and BB/V006258/1 to E.V.M.), the Wellcome Trust (Tech Dev. WT093389 and Investigator Award WT220861/Z to C.H.), and a University of Bath start-up package awarded to N.N. C.H. conceived the research project and secured funding. N.N. P.M.G. F.H. R.T. E.V.M. and C.H. designed the experiments. N.N. P.M.G. R.T. and J.L.-J. conducted the experiments. N.N. analyzed the data. F.H. performed the bioinformatic analysis of RNA-seq data. N.N. wrote the manuscript with input from all authors. The authors declare no competing interests

Publisher Copyright:
© 2022 The Author(s)

Keywords

Z+agrn
alternative splicing
mRNA processing
mRNA stability
mRNA transport
motor neurons
neurodegenerative diseases
neuromuscular junction
ribonucleoprotein complexes
synaptic connectivity

ASJC Scopus subject areas

General Neuroscience
General Biochemistry,Genetics and Molecular Biology
General Agricultural and Biological Sciences

Access to Document

10.1016/j.cub.2022.10.048Licence: CC BY

Cite this

@article{425e35e4e3d04e0eabdad0df00bd5b14,

title = "Cytoplasmic pool of U1 spliceosome protein SNRNP70 shapes the axonal transcriptome and regulates motor connectivity",

abstract = "Regulation of pre-mRNA splicing and polyadenylation plays a profound role in neurons by diversifying the proteome and modulating gene expression in response to physiological cues. Although most of the pre-mRNA processing is thought to occur in the nucleus, numerous splicing regulators are also found in neurites. Here, we show that U1-70K/SNRNP70, a component of the major spliceosome, localizes in RNA-associated granules in zebrafish axons. We identify the extra-nuclear SNRNP70 as an important regulator of motor axonal growth, nerve-dependent acetylcholine receptor (AChR) clustering, and neuromuscular synaptogenesis. This cytoplasmic pool has a protective role for a limited number of transcripts regulating their abundance and trafficking inside axons. Moreover, non-nuclear SNRNP70 regulates splice variants of transcripts such as agrin, thereby controlling synapse formation. Our results point to an unexpected, yet essential, function of non-nuclear SNRNP70 in axonal development, indicating a role of spliceosome proteins in cytoplasmic RNA metabolism during neuronal connectivity.",

keywords = "Z+agrn, alternative splicing, mRNA processing, mRNA stability, mRNA transport, motor neurons, neurodegenerative diseases, neuromuscular junction, ribonucleoprotein complexes, synaptic connectivity",

author = "Nikolas Nikolaou and Patricia Gordon and Fursham Hamid and Richard Taylor and Joshua Lloyd-Jones and Eugene Makeyev and Corinne Houart",

note = "Funding Information: We thank Prof. David Stanek for kindly giving us the human SNRNP70 construct, Prof. Martin Meyer for the HuC:Gal4 construct, Triona Fielding for technical support throughout the work, and Dr. Rachel Moore and Prof. Jon Clarke for assistance in using the Zeiss Airyscan confocal imaging system. We also thank the staff at King{\textquoteright}s College London (Guy{\textquoteright}s Campus) and the University of Bath fish facilities for their fish husbandry and care. This study was supported by the Biotechnology and Biological Sciences Research Council ( BB/P001599/1 to C.H. and BB/M007103/1 , BB/R001049/1 , and BB/V006258/1 to E.V.M.), the Wellcome Trust (Tech Dev. WT093389 and Investigator Award WT220861/Z to C.H.), and a University of Bath start-up package awarded to N.N. Funding Information: We thank Prof. David Stanek for kindly giving us the human SNRNP70 construct, Prof. Martin Meyer for the HuC:Gal4 construct, Triona Fielding for technical support throughout the work, and Dr. Rachel Moore and Prof. Jon Clarke for assistance in using the Zeiss Airyscan confocal imaging system. We also thank the staff at King's College London (Guy's Campus) and the University of Bath fish facilities for their fish husbandry and care. This study was supported by the Biotechnology and Biological Sciences Research Council (BB/P001599/1 to C.H. and BB/M007103/1, BB/R001049/1, and BB/V006258/1 to E.V.M.), the Wellcome Trust (Tech Dev. WT093389 and Investigator Award WT220861/Z to C.H.), and a University of Bath start-up package awarded to N.N. C.H. conceived the research project and secured funding. N.N. P.M.G. F.H. R.T. E.V.M. and C.H. designed the experiments. N.N. P.M.G. R.T. and J.L.-J. conducted the experiments. N.N. analyzed the data. F.H. performed the bioinformatic analysis of RNA-seq data. N.N. wrote the manuscript with input from all authors. The authors declare no competing interests Publisher Copyright: {\textcopyright} 2022 The Author(s)",

year = "2022",

month = dec,

day = "5",

doi = "10.1016/j.cub.2022.10.048",

language = "English",

volume = "32",

pages = "5099--5115.e8",

journal = "Current Biology ",

issn = "0960-9822",

publisher = "Cell Press",

number = "23",

}

TY - JOUR

T1 - Cytoplasmic pool of U1 spliceosome protein SNRNP70 shapes the axonal transcriptome and regulates motor connectivity

AU - Nikolaou, Nikolas

AU - Gordon, Patricia

AU - Hamid, Fursham

AU - Taylor, Richard

AU - Lloyd-Jones, Joshua

AU - Makeyev, Eugene

AU - Houart, Corinne

N1 - Funding Information: We thank Prof. David Stanek for kindly giving us the human SNRNP70 construct, Prof. Martin Meyer for the HuC:Gal4 construct, Triona Fielding for technical support throughout the work, and Dr. Rachel Moore and Prof. Jon Clarke for assistance in using the Zeiss Airyscan confocal imaging system. We also thank the staff at King’s College London (Guy’s Campus) and the University of Bath fish facilities for their fish husbandry and care. This study was supported by the Biotechnology and Biological Sciences Research Council ( BB/P001599/1 to C.H. and BB/M007103/1 , BB/R001049/1 , and BB/V006258/1 to E.V.M.), the Wellcome Trust (Tech Dev. WT093389 and Investigator Award WT220861/Z to C.H.), and a University of Bath start-up package awarded to N.N. Funding Information: We thank Prof. David Stanek for kindly giving us the human SNRNP70 construct, Prof. Martin Meyer for the HuC:Gal4 construct, Triona Fielding for technical support throughout the work, and Dr. Rachel Moore and Prof. Jon Clarke for assistance in using the Zeiss Airyscan confocal imaging system. We also thank the staff at King's College London (Guy's Campus) and the University of Bath fish facilities for their fish husbandry and care. This study was supported by the Biotechnology and Biological Sciences Research Council (BB/P001599/1 to C.H. and BB/M007103/1, BB/R001049/1, and BB/V006258/1 to E.V.M.), the Wellcome Trust (Tech Dev. WT093389 and Investigator Award WT220861/Z to C.H.), and a University of Bath start-up package awarded to N.N. C.H. conceived the research project and secured funding. N.N. P.M.G. F.H. R.T. E.V.M. and C.H. designed the experiments. N.N. P.M.G. R.T. and J.L.-J. conducted the experiments. N.N. analyzed the data. F.H. performed the bioinformatic analysis of RNA-seq data. N.N. wrote the manuscript with input from all authors. The authors declare no competing interests Publisher Copyright: © 2022 The Author(s)

PY - 2022/12/5

Y1 - 2022/12/5

N2 - Regulation of pre-mRNA splicing and polyadenylation plays a profound role in neurons by diversifying the proteome and modulating gene expression in response to physiological cues. Although most of the pre-mRNA processing is thought to occur in the nucleus, numerous splicing regulators are also found in neurites. Here, we show that U1-70K/SNRNP70, a component of the major spliceosome, localizes in RNA-associated granules in zebrafish axons. We identify the extra-nuclear SNRNP70 as an important regulator of motor axonal growth, nerve-dependent acetylcholine receptor (AChR) clustering, and neuromuscular synaptogenesis. This cytoplasmic pool has a protective role for a limited number of transcripts regulating their abundance and trafficking inside axons. Moreover, non-nuclear SNRNP70 regulates splice variants of transcripts such as agrin, thereby controlling synapse formation. Our results point to an unexpected, yet essential, function of non-nuclear SNRNP70 in axonal development, indicating a role of spliceosome proteins in cytoplasmic RNA metabolism during neuronal connectivity.

AB - Regulation of pre-mRNA splicing and polyadenylation plays a profound role in neurons by diversifying the proteome and modulating gene expression in response to physiological cues. Although most of the pre-mRNA processing is thought to occur in the nucleus, numerous splicing regulators are also found in neurites. Here, we show that U1-70K/SNRNP70, a component of the major spliceosome, localizes in RNA-associated granules in zebrafish axons. We identify the extra-nuclear SNRNP70 as an important regulator of motor axonal growth, nerve-dependent acetylcholine receptor (AChR) clustering, and neuromuscular synaptogenesis. This cytoplasmic pool has a protective role for a limited number of transcripts regulating their abundance and trafficking inside axons. Moreover, non-nuclear SNRNP70 regulates splice variants of transcripts such as agrin, thereby controlling synapse formation. Our results point to an unexpected, yet essential, function of non-nuclear SNRNP70 in axonal development, indicating a role of spliceosome proteins in cytoplasmic RNA metabolism during neuronal connectivity.

KW - Z+agrn

KW - alternative splicing

KW - mRNA processing

KW - mRNA stability

KW - mRNA transport

KW - motor neurons

KW - neurodegenerative diseases

KW - neuromuscular junction

KW - ribonucleoprotein complexes

KW - synaptic connectivity

UR - http://www.scopus.com/inward/record.url?scp=85143158917&partnerID=8YFLogxK

U2 - 10.1016/j.cub.2022.10.048

DO - 10.1016/j.cub.2022.10.048

M3 - Article

SN - 0960-9822

VL - 32

SP - 5099-5115.e8

JO - Current Biology

JF - Current Biology

IS - 23

ER -

Cytoplasmic pool of U1 spliceosome protein SNRNP70 shapes the axonal transcriptome and regulates motor connectivity

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Confocal Laser Scanning Confocal Microscope (LSM)

Confocal Laser Scanning Microscope Zeiss LSM-880 with Airyscan and multiphoton laser

Cite this

Cytoplasmic pool of U1 spliceosome protein SNRNP70 shapes the axonal transcriptome and regulates motor connectivity

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Equipment

Confocal Laser Scanning Confocal Microscope (LSM)

Confocal Laser Scanning Microscope Zeiss LSM-880 with Airyscan and multiphoton laser

Cite this