Coilin phosphorylation mediates interaction with SMN and SmB'.
Cajal bodies (CBs) are subnuclear domains that participate in spliceosomal small nuclear ribonucleoprotein (snRNP) biogenesis and play a part in the assembly of the spliceosomal complex. The CB marker protein, coilin, interacts with survival of motor neuron (SMN) and Sm proteins. Several coilin phosphoresidues have been identified by mass spectrometric analysis. Phosphorylation of coilin affects its self-interaction and localization in the nucleus. We hypothesize that coilin phosphorylation also impacts its binding to SMN and Sm proteins. In vitro binding studies with a C-terminal fragment of coilin and corresponding phosphomimics show that SMN binds preferentially to dephosphorylated analogs and that SmB' binds preferentially to phosphomimetic constructs. Bacterially expressed full-length coilin binds more SMN and SmB' than does the C-terminal fragment. Co-immunoprecipitation and phosphatase experiments show that SMN also binds dephosphorylated coilin in vivo. These data show that phosphorylation of coilin influences interaction with its target proteins and, thus, may be significant in managing the flow of snRNPs through the CB.
Toyota
Cory G
CG
Department of Biochemistry, University of Mississippi Medical Center, Jackson, MS 39216, USA.
Davis
Misty D
MD
Cosman
Angela M
AM
Hebert
Michael D
MD
D019943:Amino Acid Substitution; D002460:Cell Line; D020541:Coiled Bodies / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D055533:Survival of Motor Neuron 1 Protein / Q000737:chemistry / Q000378:metabolism*; D055517:snRNP Core Proteins / Q000737:chemistry / Q000378:metabolism*
D009687:Nuclear Proteins; D017411:Ribonucleoproteins, Small Nuclear; C526407:SNRPB protein, human; D055533:Survival of Motor Neuron 1 Protein; D055517:snRNP Core Proteins; C068921:p80-coilin