Ectopic expression of the Aspergillus nidulans mitotic inducer, nimA kinase, in megakaryocytes: effect on polyploidization.
Aspergillus nidulans nimA gene encodes a serine/threonine protein kinase (NIMA) whose activity is essential for mitotic entry and chromatin condensation. Both the activity and the abundance of NIMA protein increase at the G2/M transition of the fungal cell cycle. In this study, we report the effects elicited by ectopic expression of nimA on polyploidization in a mouse megakaryocytic line, Y10, which is undergoing an endomitotic cell cycle. A pool of Y10 stable transfectants that have been induced to express nimA displayed a decrease in cell number and an elevated DNA content per cell. NIMA also dramatically enhanced the activity of phorbal 12-myristate 13-acetate toward polyploidization. Analysis of individual nimA transfectants revealed that the DNA content per cell rose in cells expressing high levels of nimA and that the level of cyclin B was reduced as compared to the mock-transfected cells. These effects observed in polyploidizing megakaryocytes are in contrast to those found in A. nidulans and HeLa cells, in which induced nimA expression caused abnormal chromatin condensation and cell cycle arrest. We conclude that high-level expression of nimA in cells programmed to undergo endomitosis could potentiate polyploidization. The challenge now resides in the isolation of the authentic megakaryocyte counterpart of the fungal nimA.
Sun
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Department of Biochemistry and Whitaker Cardiovascular Institute, Boston University School of Medicine, MA 02118, USA.
Kaluzhny
Y
Y
Ravid
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K
D000818:Animals; D001233:Aspergillus nidulans / Q000201:enzymology* / Q000235:genetics; D018797:Cell Cycle Proteins; D002460:Cell Line; D002843:Chromatin; D019926:Cyclin B / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D008533:Megakaryocytes / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008938:Mitosis; D000072380:NIMA-Related Kinase 1; D000072379:NIMA-Related Kinases; D011123:Polyploidy; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D018797:Cell Cycle Proteins; D002843:Chromatin; D019926:Cyclin B; D004247:DNA; D000072380:NIMA-Related Kinase 1; D000072379:NIMA-Related Kinases; C442331:NIMA-related kinase 6; C000606920:Nek1 protein, mouse; D017346:Protein Serine-Threonine Kinases; D013755:Tetradecanoylphorbol Acetate