A cell system with targeted disruption of the SMN gene: functional conservation of the SMN protein and dependence of Gemin2 on SMN - PubMed (original) (raw)
. 2001 Mar 30;276(13):9599-605.
doi: 10.1074/jbc.M009162200. Epub 2000 Dec 19.
Affiliations
- PMID: 11121410
- DOI: 10.1074/jbc.M009162200
Free article
A cell system with targeted disruption of the SMN gene: functional conservation of the SMN protein and dependence of Gemin2 on SMN
J Wang et al. J Biol Chem. 2001.
Free article
Abstract
The motor neuron degenerative disease spinal muscular atrophy is caused by reduced expression of the survival motor neuron (SMN) protein. Here we report a genetic system developed in the chicken pre-B cell line DT40, in which the endogenous SMN gene is disrupted by homologous recombination, and SMN protein is expressed from a chicken SMN cDNA under control of a tetracycline (tet)-repressible promoter. Addition of tet results in depletion of SMN protein and consequent cell death, which directly demonstrates that SMN is required for cell viability. The tet-induced lethality can be rescued by expression of human SMN, indicating that the function of SMN is highly conserved between the two species. Cells expressing low levels of SMN display slow growth proportional to the amount of SMN they contain. Interestingly, the level of the SMN-interacting protein Gemin2 decreases significantly following depletion of SMN, supporting the conclusion that SMN and Gemin2 form a stable complex in vivo. This system provides a powerful setting for studying the function of SMN in vivo and for screening for potential therapeutics for spinal muscular atrophy.
Similar articles
- Characterization of functional domains of the SMN protein in vivo.
Wang J, Dreyfuss G. Wang J, et al. J Biol Chem. 2001 Nov 30;276(48):45387-93. doi: 10.1074/jbc.M105059200. Epub 2001 Sep 25. J Biol Chem. 2001. PMID: 11572858 - Identification of a novel cyclic AMP-response element (CRE-II) and the role of CREB-1 in the cAMP-induced expression of the survival motor neuron (SMN) gene.
Majumder S, Varadharaj S, Ghoshal K, Monani U, Burghes AH, Jacob ST. Majumder S, et al. J Biol Chem. 2004 Apr 9;279(15):14803-11. doi: 10.1074/jbc.M308225200. Epub 2004 Jan 23. J Biol Chem. 2004. PMID: 14742439 Free PMC article. Retracted. - A transgene carrying an A2G missense mutation in the SMN gene modulates phenotypic severity in mice with severe (type I) spinal muscular atrophy.
Monani UR, Pastore MT, Gavrilina TO, Jablonka S, Le TT, Andreassi C, DiCocco JM, Lorson C, Androphy EJ, Sendtner M, Podell M, Burghes AH. Monani UR, et al. J Cell Biol. 2003 Jan 6;160(1):41-52. doi: 10.1083/jcb.200208079. Epub 2003 Jan 6. J Cell Biol. 2003. PMID: 12515823 Free PMC article. - Fishing for a mechanism: using zebrafish to understand spinal muscular atrophy.
Beattie CE, Carrel TL, McWhorter ML. Beattie CE, et al. J Child Neurol. 2007 Aug;22(8):995-1003. doi: 10.1177/0883073807305671. J Child Neurol. 2007. PMID: 17761655 Review. - Therapeutics development for spinal muscular atrophy.
Sumner CJ. Sumner CJ. NeuroRx. 2006 Apr;3(2):235-45. doi: 10.1016/j.nurx.2006.01.010. NeuroRx. 2006. PMID: 16554261 Free PMC article. Review.
Cited by
- Splicing efficiency of minor introns in a mouse model of SMA predominantly depends on their branchpoint sequence and can involve the contribution of major spliceosome components.
Jacquier V, Prévot M, Gostan T, Bordonné R, Benkhelifa-Ziyyat S, Barkats M, Soret J. Jacquier V, et al. RNA. 2022 Mar;28(3):303-319. doi: 10.1261/rna.078329.120. Epub 2021 Dec 10. RNA. 2022. PMID: 34893560 Free PMC article. - What Genetics Has Told Us and How It Can Inform Future Experiments for Spinal Muscular Atrophy, a Perspective.
Blatnik AJ 3rd, McGovern VL, Burghes AHM. Blatnik AJ 3rd, et al. Int J Mol Sci. 2021 Aug 6;22(16):8494. doi: 10.3390/ijms22168494. Int J Mol Sci. 2021. PMID: 34445199 Free PMC article. - Conditional deletion of SMN in cell culture identifies functional SMN alleles.
Blatnik AJ, McGovern VL, Le TT, Iyer CC, Kaspar BK, Burghes AHM. Blatnik AJ, et al. Hum Mol Genet. 2020 Nov 1;29(21):3477-3492. doi: 10.1093/hmg/ddaa229. Epub 2020 Oct 19. Hum Mol Genet. 2020. PMID: 33075805 Free PMC article. - Composition of the Survival Motor Neuron (SMN) Complex in Drosophila melanogaster.
Matera AG, Raimer AC, Schmidt CA, Kelly JA, Droby GN, Baillat D, Ten Have S, Lamond AI, Wagner EJ, Gray KM. Matera AG, et al. G3 (Bethesda). 2019 Feb 7;9(2):491-503. doi: 10.1534/g3.118.200874. G3 (Bethesda). 2019. PMID: 30563832 Free PMC article. - The role of survival motor neuron protein (SMN) in protein homeostasis.
Chaytow H, Huang YT, Gillingwater TH, Faller KME. Chaytow H, et al. Cell Mol Life Sci. 2018 Nov;75(21):3877-3894. doi: 10.1007/s00018-018-2849-1. Epub 2018 Jun 5. Cell Mol Life Sci. 2018. PMID: 29872871 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources