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PMID: 11470819 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Residual Cajal bodies in coilin knockout mice fail to recruit Sm snRNPs and SMN, the spinal muscular atrophy gene product.

The Journal of cell biology ·Vol. 154 ·No. 2 ·2001-07-23 ·Pages 293-307

Tucker KE, Berciano MT, Jacobs EY, LePage DF, Shpargel KB, Rossire JJ, Chan EK, Lafarga M, Conlon RA, Matera AG

Abstract

Cajal bodies (CBs) are nuclear suborganelles involved in the biogenesis of small nuclear ribonucleoproteins (snRNPs). In addition to snRNPs, they are highly enriched in basal transcription and cell cycle factors, the nucleolar proteins fibrillarin (Fb) and Nopp140 (Nopp), the survival motor neuron (SMN) protein complex, and the CB marker protein, p80 coilin. We report the generation of knockout mice lacking the COOH-terminal 487 amino acids of coilin. Northern and Western blot analyses demonstrate that we have successfully removed the full-length coilin protein from the knockout animals. Some homozygous mutant animals are viable, but their numbers are reduced significantly when crossed to inbred backgrounds. Analysis of tissues and cell lines from mutant animals reveals the presence of extranucleolar foci that contain Fb and Nopp but not other typical nucleolar markers. These so-called "residual" CBs neither condense Sm proteins nor recruit members of the SMN protein complex. Transient expression of wild-type mouse coilin in knockout cells results in formation of CBs and restores these missing epitopes. Our data demonstrate that full-length coilin is essential for proper formation and/or maintenance of CBs and that recruitment of snRNP and SMN complex proteins to these nuclear subdomains requires sequences within the coilin COOH terminus.

MeSH Terms
Animals Autoantigens/metabolism Blotting, Northern Cell Line Cell Nucleolus/metabolism,ultrastructure Chromosomal Proteins, Non-Histone/metabolism Coiled Bodies/genetics,metabolism,ultrastructure Cyclic AMP Response Element-Binding Protein Fetal Viability/genetics Gene Expression/drug effects Gene Targeting Green Fluorescent Proteins Homozygote Luminescent Proteins/genetics Mice Mice, Knockout/genetics Nerve Tissue Proteins/metabolism Nuclear Proteins/biosynthesis,deficiency,genetics,metabolism Organ Specificity Phosphoproteins/metabolism RNA Splicing RNA, Messenger RNA-Binding Proteins Recombinant Fusion Proteins/biosynthesis,genetics,pharmacology Ribonucleoproteins, Small Nuclear/metabolism SMN Complex Proteins Survival Rate snRNP Core Proteins
Chemicals
Autoantigens Chromosomal Proteins, Non-Histone Cyclic AMP Response Element-Binding Protein Luminescent Proteins NOLC1 protein, human Nerve Tissue Proteins Nolc1 protein, mouse Nuclear Proteins Phosphoproteins RNA, Messenger RNA-Binding Proteins Recombinant Fusion Proteins Ribonucleoproteins, Small Nuclear SMN Complex Proteins fibrillarin snRNP Core Proteins p80-coilin Green Fluorescent Proteins
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Tucker K E
Department of Genetics, and Program in Cell Biology, Case Western Reserve University and University Hospitals of Cleveland, Cleveland, OH 44106, USA.
Berciano M T
Jacobs E Y
LePage D F
Shpargel K B
Rossire J J
Chan E K
Lafarga M
Conlon R A
Matera A G
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200104083
Published
2001-07-23
Pages
293-307
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2150753
Subset
IM
Grants
NIGMS NIH HHS · R01 GM053034 · United States
NINDS NIH HHS · R01 NS041617 · United States
NIGMS NIH HHS · GM53034 · United States
NINDS NIH HHS · NS41617 · United States
External Links
PubMed source DOI: 10.1083/jcb.200104083 PMC Full Text View journal details
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