Tim50a, a nuclear isoform of the mitochondrial Tim50, interacts with proteins involved in snRNP biogenesis.
BACKGROUND
The Cajal body (CB) is a nuclear suborganelle involved in the biogenesis of small nuclear ribonucleoproteins (snRNPs), which are vital for pre-mRNA splicing. Newly imported Sm-class snRNPs traffic through CBs, where the snRNA component of the snRNP is modified, and then target to other nuclear domains such as speckles and perichromatin fibrils. It is not known how nascent snRNPs localize to the CB and are released from this structure after modification. The marker protein for CBs, coilin, may play a role in snRNP biogenesis given that it can interact with snRNPs and SMN, the protein mutated in Spinal Muscular Atrophy. Loss of coilin function in mice leads to significant viability and fertility problems and altered CB formation.


RESULTS
In this report, we identify a minor isoform of the mitochondrial Tim50, Tim50a, as a coilin interacting protein. The Tim50a transcript can be detected in some cancer cell lines and normal brain tissue. The Tim50a protein differs only from Tim50 in that it contains an additional 103 aa N-terminal to the translation start of Tim50. Importantly, a putative nuclear localization signal is found within these 103 residues. In contrast to Tim50, which localizes to the cytoplasm and mitochondria, Tim50a is strictly nuclear and is enriched in speckles with snRNPs. In addition to coilin, Tim50a interacts with snRNPs and SMN. Competition binding experiments demonstrate that coilin competes with Sm proteins of snRNPs and SMN for binding sites on Tim50a.


CONCLUSIONS
Tim50a may play a role in snRNP biogenesis given its cellular localization and protein interaction characteristics. We hypothesize that Tim50a takes part in the release of snRNPs and SMN from the CB.
Xu
Hongzhi
H
Department of Biochemistry, The University of Mississippi Medical Center Jackson, MS 39216-4505, USA. hxu@biochem.umsmed.edu
Somers
Z Brad
ZB
Robinson
Melvin L
ML
Hebert
Michael D
MD
D001921:Brain; D045744:Cell Line, Tumor; D006367:HeLa Cells; D006801:Humans; D026901:Membrane Transport Proteins / Q000502:physiology*; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins / Q000378:metabolism*; D020033:Protein Isoforms; D017411:Ribonucleoproteins, Small Nuclear / Q000096:biosynthesis* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D026901:Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins; D020033:Protein Isoforms; D017411:Ribonucleoproteins, Small Nuclear; C509348:TIMM50 protein, human; C068921:p80-coilin