Abstract
Aim:
RNA-binding proteins are a large group of regulators (800–1000 in humans), some of which play significant roles in mRNA local translation. In this study, we analyzed the functions of the protein RNP-1, which was previously discovered in a genetic selection screen for nocodazole suppression.
Methods:
The growth rates and the microtubule networks of Dictyostelium cells were assessed with or without nocodazole (10 μmol/L) in suspension culture. Fluorescent images of RNP-1-GFP and RFP-tubulin were captured when cells were undergoing cytokinesis, then the GFP signal intensity and distance to the nearest centrosome were analyzed by using a computer program written in Matlab®. The RNP-1-GFP-expresseding cells were polarized, and the time-lapse images of cells were captured when cells were chemotaxing to a cAMP source.
Results:
Over-expression of RNP-1 rescued the growth defects caused by the microtubule-destabilizing agent nocodazole. Over-expression of RNP-1 protected microtubules from nocodazole treatment. In cells undergoing cytokinesis, the RNP-1 protein was localized to the polar regions of the cell cortex, and protein levels decreased proportionally as the power of the distance from the cell cortex to the nearest centrosome. In chemotactic cells, the RNP-1 protein localized to the leading edge of moving cells. Sequence analysis revealed that RNP-1 has two RNA-binding domains and is related to cytosolic poly(A)-binding proteins (PABPCs) in humans.
Conclusion:
RNP-1 has roles in protecting microtubules and in directing cortical movement during cytokinesis and cell migration in Dictyostelium cells. The sequence similarity of RNP-1 to human PABPCs suggests that PABPCs may have similar functions in mammalian cells, perhaps in regulating microtubule dynamics and functions during cortical movement in cytokinesis and cell migration.
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Acknowledgements
The majority of the bench experiments were performed in the laboratory of Dr Douglas N ROBINSON, and we thank him for his generous support. We also thank the NIH (R01 GM66817 to Douglas N ROBINSON) and AHA (Qiong-qiong ZHOU's fellowship) for support.
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Supplementary information is available on the website of Acta Pharmacologica Sinica.
Supplementary information
Supplementary Movie S1
GFP-tubulin labeled 3D microtubule network in a Dictyostelium cell treated with 10 μmol/L of DMSO. (AVI 321 kb)
Supplementary Movie S2
GFP-tubulin labeled 3D microtubule network in a Dictyostelium cell treated with 10 μmol/L of nocodazole. (AVI 456 kb)
Supplementary Movie S3
RNP-1-GFP localization in Dictyostelium cells during chemotaxing toward cAMP needles. (AVI 171 kb)
Supplementary Movie S4
RNP-1-GFP localization in Dictyostelium cells during chemotaxing toward cAMP needles. (AVI 168 kb)
Supplementary Figure S1
Microtubule networks in the surface cultured cells after overnight nacodazole treatment. (JPG 1135 kb)
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Ngo, T., Miao, X., Robinson, D. et al. An RNA-binding protein, RNP-1, protects microtubules from nocodazole and localizes to the leading edge during cytokinesis and cell migration in Dictyostelium cells. Acta Pharmacol Sin 37, 1449–1457 (2016). https://doi.org/10.1038/aps.2016.57
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DOI: https://doi.org/10.1038/aps.2016.57
