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© The Rockefeller University Press, 0021-9525/1997//871 $5.00
The Journal of Cell Biology, Volume 137, Number 4, , 1997 871-879

Article

mRNAs for Microtubule Proteins Are Specifically Colocalized during the Sequential Formation of Basal Body, Flagella, and Cytoskeletal Microtubules in the Differentiation of Naegleria gruberi



Ji Woong Han, Jong Ho Park, Misook Kim, and JooHun Lee

Department of Biology, Yonsei University, Seoul, Korea 120-749

We have examined the distribution of four mRNAs—{alpha}-tubulin, β-tubulin, flagellar calmodulin, and Class I mRNA—during differentiation of Naegleria gruberi amebas into flagellates by in situ hybridization. Three of the four mRNAs—{alpha}-tubulin, β-tubulin, and Class I mRNA—began to be colocalized at the periphery of the cells as soon as transcription of the respective genes was activated and before any microtubular structures were observable. At 70 min after the initiation of differentiation, these mRNAs were relocalized to the base of the growing flagella, adjacent to the basal bodies and microtubule organizing center for the cytoskeletal microtubules. Within an additional 15 min, the mRNAs were translocated to the posterior of the flagellated cells, and by the end of differentiation (120 min), very low levels of the mRNAs were observed. Cytochalasin D inhibited stage-specific localization of the mRNAs, demonstrating that RNA localization was actin dependent. Since cytochalasin D also blocked differentiation, this raises the possibility that actin-dependent RNA movement is an essential process for differentiation.

1. Abbreviations used in this paper: CSMT, cytoskeletal microtubules; DIC, differential interference contrast; DIG, digoxygenin; DS, differentiation-specific.

This work is supported by grants from Korea Science and Engineering Foundation (94-0401-08-04-03) and Yonsei University (1993, 1995).

Address all correspondence to JooHun Lee, Department of Biology, Yonsei University, 134 ShinChon-Dong, SeoDaiMoon-Gu, Seoul, Korea 120-749. Tel.: 82-2-361-2659. Fax: 82-2-312-5657. E-mail: leehjnt{at}bubble.yonsei.ac.kr


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