Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster

A. Gregory Matera (Lead / Corresponding author), Amanda C. Raimer, Casey A. Schmidt, Jo A. Kelly, Gaith N. Droby, David Baillat, Sara Ten Have, Angus I. Lamond, Eric J. Wagner, Kelsey M. Gray

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Abstract

Spinal Muscular Atrophy (SMA) is caused by homozygous mutations in the human survival motor neuron 1 (SMN1) gene. SMN protein has a well-characterized role in the biogenesis of small nuclear ribonucleoproteins (snRNPs), core components of the spliceosome. SMN is part of an oligomeric complex with core binding partners, collectively called Gemins. Biochemical and cell biological studies demonstrate that certain Gemins are required for proper snRNP assembly and transport. However, the precise functions of most Gemins are unknown. To gain a deeper understanding of the SMN complex in the context of metazoan evolution, we investigated the composition of the SMN complex in Drosophila melanogaster. Using a stable transgenic line that exclusively expresses Flag-tagged SMN from its native promoter, we previously found that Gemin2, Gemin3, Gemin5, and all nine classical Sm proteins, including Lsm10 and Lsm11, co-purify with SMN. Here, we show that CG2941 is also highly enriched in the pulldown. Reciprocal co-immunoprecipitation reveals that epitope-tagged CG2941 interacts with endogenous SMN in Schneider2 cells. Bioinformatic comparisons show that CG2941 shares sequence and structural similarity with metazoan Gemin4. Additional analysis shows that three other genes (CG14164, CG31950 and CG2371) are not orthologous to Gemins 6-7-8, respectively, as previously suggested. In D. melanogaster, CG2941 is located within an evolutionarily recent genomic triplication with two other nearly identical paralogous genes (CG32783 and CG32786). RNAi-mediated knockdown of CG2941 and its two close paralogs reveals that Gemin4 is essential for organismal viability.

Original languageEnglish
Pages (from-to)491-503
Number of pages13
JournalG3 : Genes, Genomes, Genetics
Volume9
Issue number2
Early online date21 Dec 2018
DOIs
Publication statusPublished - 1 Feb 2019

Fingerprint

SMN Complex Proteins
Motor Neurons
Drosophila melanogaster
Survival
Small Nuclear Ribonucleoproteins
Spliceosomes
Genes
Spinal Muscular Atrophy
RNA Interference
Computational Biology
Immunoprecipitation
Epitopes
Proteins
Mutation

Keywords

  • Atrophy
  • Function
  • Locomotor
  • Neuron
  • Proteomics
  • RNP assembly
  • SMA
  • SMN
  • Spinal Muscular
  • Survival motor
  • ncRNA
  • snRNA
  • snRNP

Cite this

Matera, A. G., Raimer, A. C., Schmidt, C. A., Kelly, J. A., Droby, G. N., Baillat, D., ... Gray, K. M. (2019). Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster. G3 : Genes, Genomes, Genetics, 9(2), 491-503. https://doi.org/10.1534/g3.118.200874
Matera, A. Gregory ; Raimer, Amanda C. ; Schmidt, Casey A. ; Kelly, Jo A. ; Droby, Gaith N. ; Baillat, David ; Ten Have, Sara ; Lamond, Angus I. ; Wagner, Eric J. ; Gray, Kelsey M. / Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster. In: G3 : Genes, Genomes, Genetics. 2019 ; Vol. 9, No. 2. pp. 491-503.
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Matera, AG, Raimer, AC, Schmidt, CA, Kelly, JA, Droby, GN, Baillat, D, Ten Have, S, Lamond, AI, Wagner, EJ & Gray, KM 2019, 'Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster', G3 : Genes, Genomes, Genetics, vol. 9, no. 2, pp. 491-503. https://doi.org/10.1534/g3.118.200874

Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster. / Matera, A. Gregory (Lead / Corresponding author); Raimer, Amanda C.; Schmidt, Casey A. ; Kelly, Jo A. ; Droby, Gaith N. ; Baillat, David; Ten Have, Sara; Lamond, Angus I.; Wagner, Eric J.; Gray, Kelsey M.

In: G3 : Genes, Genomes, Genetics, Vol. 9, No. 2, 01.02.2019, p. 491-503.

Research output: Contribution to journalArticle

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T1 - Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster

AU - Matera, A. Gregory

AU - Raimer, Amanda C.

AU - Schmidt, Casey A.

AU - Kelly, Jo A.

AU - Droby, Gaith N.

AU - Baillat, David

AU - Ten Have, Sara

AU - Lamond, Angus I.

AU - Wagner, Eric J.

AU - Gray, Kelsey M.

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N2 - Spinal Muscular Atrophy (SMA) is caused by homozygous mutations in the human survival motor neuron 1 (SMN1) gene. SMN protein has a well-characterized role in the biogenesis of small nuclear ribonucleoproteins (snRNPs), core components of the spliceosome. SMN is part of an oligomeric complex with core binding partners, collectively called Gemins. Biochemical and cell biological studies demonstrate that certain Gemins are required for proper snRNP assembly and transport. However, the precise functions of most Gemins are unknown. To gain a deeper understanding of the SMN complex in the context of metazoan evolution, we investigated the composition of the SMN complex in Drosophila melanogaster. Using a stable transgenic line that exclusively expresses Flag-tagged SMN from its native promoter, we previously found that Gemin2, Gemin3, Gemin5, and all nine classical Sm proteins, including Lsm10 and Lsm11, co-purify with SMN. Here, we show that CG2941 is also highly enriched in the pulldown. Reciprocal co-immunoprecipitation reveals that epitope-tagged CG2941 interacts with endogenous SMN in Schneider2 cells. Bioinformatic comparisons show that CG2941 shares sequence and structural similarity with metazoan Gemin4. Additional analysis shows that three other genes (CG14164, CG31950 and CG2371) are not orthologous to Gemins 6-7-8, respectively, as previously suggested. In D. melanogaster, CG2941 is located within an evolutionarily recent genomic triplication with two other nearly identical paralogous genes (CG32783 and CG32786). RNAi-mediated knockdown of CG2941 and its two close paralogs reveals that Gemin4 is essential for organismal viability.

AB - Spinal Muscular Atrophy (SMA) is caused by homozygous mutations in the human survival motor neuron 1 (SMN1) gene. SMN protein has a well-characterized role in the biogenesis of small nuclear ribonucleoproteins (snRNPs), core components of the spliceosome. SMN is part of an oligomeric complex with core binding partners, collectively called Gemins. Biochemical and cell biological studies demonstrate that certain Gemins are required for proper snRNP assembly and transport. However, the precise functions of most Gemins are unknown. To gain a deeper understanding of the SMN complex in the context of metazoan evolution, we investigated the composition of the SMN complex in Drosophila melanogaster. Using a stable transgenic line that exclusively expresses Flag-tagged SMN from its native promoter, we previously found that Gemin2, Gemin3, Gemin5, and all nine classical Sm proteins, including Lsm10 and Lsm11, co-purify with SMN. Here, we show that CG2941 is also highly enriched in the pulldown. Reciprocal co-immunoprecipitation reveals that epitope-tagged CG2941 interacts with endogenous SMN in Schneider2 cells. Bioinformatic comparisons show that CG2941 shares sequence and structural similarity with metazoan Gemin4. Additional analysis shows that three other genes (CG14164, CG31950 and CG2371) are not orthologous to Gemins 6-7-8, respectively, as previously suggested. In D. melanogaster, CG2941 is located within an evolutionarily recent genomic triplication with two other nearly identical paralogous genes (CG32783 and CG32786). RNAi-mediated knockdown of CG2941 and its two close paralogs reveals that Gemin4 is essential for organismal viability.

KW - Atrophy

KW - Function

KW - Locomotor

KW - Neuron

KW - Proteomics

KW - RNP assembly

KW - SMA

KW - SMN

KW - Spinal Muscular

KW - Survival motor

KW - ncRNA

KW - snRNA

KW - snRNP

U2 - 10.1534/g3.118.200874

DO - 10.1534/g3.118.200874

M3 - Article

VL - 9

SP - 491

EP - 503

JO - G3 : Genes, Genomes, Genetics

JF - G3 : Genes, Genomes, Genetics

SN - 2160-1836

IS - 2

ER -

Matera AG, Raimer AC, Schmidt CA, Kelly JA, Droby GN, Baillat D et al. Composition of the Survival Motor Neuron (SMN) complex in Drosophila melanogaster. G3 : Genes, Genomes, Genetics. 2019 Feb 1;9(2):491-503. https://doi.org/10.1534/g3.118.200874