A microtubule associated protein (hNUDC) binds to the extracellular domain of thrombopoietin receptor (Mpl).
Human NUDC (hNUDC) was initially characterized as a nuclear migration protein based on the similarity of its C-terminus to that of fungal NUDC from Aspergillus nidulans. However, hNUDC is a 331 amino acid protein whereas fungal NUDC is 198 amino acids in length. The extra N-terminal portion of hNUDC has no known function or homology to other proteins. In this study, we report the binding of hNUDC to the extracellular domain of the thrombopoietin receptor (Mpl) as detected by the yeast two-hybrid system, GST pull-down, and co-immunoprecipitation. Our deletion analysis demonstrated that amino acids between positions 100 and 238 as the critical domain mediating the hNUDC and Mpl interactions as detected by the two-hybrid system and GST pull-down assay. Immunofluorescence staining of human megakaryocyte cells indicated that hNUDC and Mpl colocalized at all stages of megakaryocyte development. Substantial colocalization of hNUDC with microtubules was also detected around nuclei and elongated microtubular structures, especially in proplatelet extensions.
Pan
Rui-Min
RM
Biotechnology Research Center, The Key Laboratory of Gene Engineering of Education Ministry, Zhongshan University, Guangzhou 510275, China.
Yang
Yan
Y
Wei
Ming-Xu
MX
Yu
Xiao-Bin
XB
Ge
Yi-Chen
YC
Xu
Peilin
P
D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D006801:Humans; D047468:Immunoprecipitation; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018121:Receptors, Cytokine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D053628:Receptors, Thrombopoietin; D011993:Recombinant Fusion Proteins; D017384:Sequence Deletion; D014404:Tubulin / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D018797:Cell Cycle Proteins; C512648:NUDC protein, human; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D018121:Receptors, Cytokine; D053628:Receptors, Thrombopoietin; D011993:Recombinant Fusion Proteins; D014404:Tubulin; C082218:MPL protein, human