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mRNAs for microtubule proteins are specifically colocalized during the sequential formation of basal body, flagella, and cytoskeletal microtubules in the differentiation of Naegleria gruberi.

Han JW, Park JH, Kim M, Lee J - J. Cell Biol. (1997)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Yonsei University, Seoul, Korea 120-749.

ABSTRACT
We have examined the distribution of four mRNAs-alpha-tubulin, beta-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, beta-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.

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Cytochalasin D inhibited the specific localization of the  β-tubulin mRNA. Cytochalasin D was added at the beginning of  differentiation to a final concentration of 50 μg/ml. Cells were  taken at 20 and 70 min after the initiation and fixed. The fixed  cells were in situ hybridized with DIG-labeled β-tubulin cDNA  probe, and the location of the β-tubulin mRNA was determined  as in Fig. 1. (A) 20 min; (B) 70 min. Bar, 10 μm.
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Figure 6: Cytochalasin D inhibited the specific localization of the β-tubulin mRNA. Cytochalasin D was added at the beginning of differentiation to a final concentration of 50 μg/ml. Cells were taken at 20 and 70 min after the initiation and fixed. The fixed cells were in situ hybridized with DIG-labeled β-tubulin cDNA probe, and the location of the β-tubulin mRNA was determined as in Fig. 1. (A) 20 min; (B) 70 min. Bar, 10 μm.

Mentions: Addition of cytochalasin D at the beginning of differentiation inhibited the differentiation in a dose-dependent manner. In 20 μg/ml of cytochalasin D, 60% of the cells formed flagella. In 50 or 100 μg/ml of cytochalasin D, the differentiation was strongly inhibited (Fig. 5 A). In these experiments, most of the cells changed their shape into spheres less than 30 min after the initiation of differentiation and remained as spheres until the end of the differentiation (Fig. 6).


mRNAs for microtubule proteins are specifically colocalized during the sequential formation of basal body, flagella, and cytoskeletal microtubules in the differentiation of Naegleria gruberi.

Han JW, Park JH, Kim M, Lee J - J. Cell Biol. (1997)

Cytochalasin D inhibited the specific localization of the  β-tubulin mRNA. Cytochalasin D was added at the beginning of  differentiation to a final concentration of 50 μg/ml. Cells were  taken at 20 and 70 min after the initiation and fixed. The fixed  cells were in situ hybridized with DIG-labeled β-tubulin cDNA  probe, and the location of the β-tubulin mRNA was determined  as in Fig. 1. (A) 20 min; (B) 70 min. Bar, 10 μm.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2139841&req=5

Figure 6: Cytochalasin D inhibited the specific localization of the β-tubulin mRNA. Cytochalasin D was added at the beginning of differentiation to a final concentration of 50 μg/ml. Cells were taken at 20 and 70 min after the initiation and fixed. The fixed cells were in situ hybridized with DIG-labeled β-tubulin cDNA probe, and the location of the β-tubulin mRNA was determined as in Fig. 1. (A) 20 min; (B) 70 min. Bar, 10 μm.
Mentions: Addition of cytochalasin D at the beginning of differentiation inhibited the differentiation in a dose-dependent manner. In 20 μg/ml of cytochalasin D, 60% of the cells formed flagella. In 50 or 100 μg/ml of cytochalasin D, the differentiation was strongly inhibited (Fig. 5 A). In these experiments, most of the cells changed their shape into spheres less than 30 min after the initiation of differentiation and remained as spheres until the end of the differentiation (Fig. 6).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Yonsei University, Seoul, Korea 120-749.

ABSTRACT
We have examined the distribution of four mRNAs-alpha-tubulin, beta-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, beta-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.

Show MeSH
Related in: MedlinePlus