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Tea2p is a kinesin-like protein required to generate polarized growth in fission yeast.

Browning H, Hayles J, Mata J, Aveline L, Nurse P, McIntosh JR - J. Cell Biol. (2000)

Bottom Line: They have a reduced cytoplasmic microtubule network and display severe morphological defects in genetic backgrounds that produce long cells.The tip-specific marker, Tea1p, is mislocalized in both tea2-1 and tea2Delta cells, indicating that Tea2p function is necessary for proper localization of Tea1p.These results suggest that this kinesin promotes microtubule growth, possibly through interactions with the microtubule end, and that it is important for establishing and maintaining polarized growth along the long axis of the cell.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347, USA. browninh@icrf.icnet.uk

ABSTRACT
Cytoplasmic microtubules are critical for establishing and maintaining cell shape and polarity. Our investigations of kinesin-like proteins (klps) and morphological mutants in the fission yeast Schizosaccharomyces pombe have identified a kinesin-like gene, tea2(+), that is required for cells to generate proper polarized growth. Cells deleted for this gene are often bent during exponential growth and initiate growth from improper sites as they exit stationary phase. They have a reduced cytoplasmic microtubule network and display severe morphological defects in genetic backgrounds that produce long cells. The tip-specific marker, Tea1p, is mislocalized in both tea2-1 and tea2Delta cells, indicating that Tea2p function is necessary for proper localization of Tea1p. Tea2p is localized to the tips of the cell and in a punctate pattern within the cell, often coincident with the ends of cytoplasmic microtubules. These results suggest that this kinesin promotes microtubule growth, possibly through interactions with the microtubule end, and that it is important for establishing and maintaining polarized growth along the long axis of the cell.

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DIC microscopy of (A) wild-type and (B) tea2Δ cells grown to mid-log phase. DIC microscopy of (C) wild-type, (D) tea2Δ, (E) cdc25-22, and (F) tea2Δ cdc25-22 cells from colonies on plates and (G) tea2Δ diploid cells from liquid culture. Bar: 10 μm.
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Figure 2: DIC microscopy of (A) wild-type and (B) tea2Δ cells grown to mid-log phase. DIC microscopy of (C) wild-type, (D) tea2Δ, (E) cdc25-22, and (F) tea2Δ cdc25-22 cells from colonies on plates and (G) tea2Δ diploid cells from liquid culture. Bar: 10 μm.

Mentions: To investigate further the cellular roles of Tea2p in S. pombe, a deletion allele was constructed by replacing the tea2+ ORF with his3+. Transformants were screened by PCR, and homologous integration was confirmed by Southern blot analysis (not shown). At 32°C, tea2Δ cells grow at rates similar to wild-type cells. These cells were examined by DIC microscopy to see if the deletion had an effect on the morphology of the cells. Cultures of exponentially growing cells contain ∼18% (n = 117) obviously bent cells, whereas wild-type cells were generally straight cylinders, 0% bent (n = 138; Fig. 2A and Fig. B). At 37°C, tea2Δ cells grew more slowly than wild-type cells, and a high percentage of T shaped cells were seen in the culture (up to 9%). These defects in cell morphology are similar to the tea2-1 mutant (Verde et al. 1995).


Tea2p is a kinesin-like protein required to generate polarized growth in fission yeast.

Browning H, Hayles J, Mata J, Aveline L, Nurse P, McIntosh JR - J. Cell Biol. (2000)

DIC microscopy of (A) wild-type and (B) tea2Δ cells grown to mid-log phase. DIC microscopy of (C) wild-type, (D) tea2Δ, (E) cdc25-22, and (F) tea2Δ cdc25-22 cells from colonies on plates and (G) tea2Δ diploid cells from liquid culture. Bar: 10 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2189814&req=5

Figure 2: DIC microscopy of (A) wild-type and (B) tea2Δ cells grown to mid-log phase. DIC microscopy of (C) wild-type, (D) tea2Δ, (E) cdc25-22, and (F) tea2Δ cdc25-22 cells from colonies on plates and (G) tea2Δ diploid cells from liquid culture. Bar: 10 μm.
Mentions: To investigate further the cellular roles of Tea2p in S. pombe, a deletion allele was constructed by replacing the tea2+ ORF with his3+. Transformants were screened by PCR, and homologous integration was confirmed by Southern blot analysis (not shown). At 32°C, tea2Δ cells grow at rates similar to wild-type cells. These cells were examined by DIC microscopy to see if the deletion had an effect on the morphology of the cells. Cultures of exponentially growing cells contain ∼18% (n = 117) obviously bent cells, whereas wild-type cells were generally straight cylinders, 0% bent (n = 138; Fig. 2A and Fig. B). At 37°C, tea2Δ cells grew more slowly than wild-type cells, and a high percentage of T shaped cells were seen in the culture (up to 9%). These defects in cell morphology are similar to the tea2-1 mutant (Verde et al. 1995).

Bottom Line: They have a reduced cytoplasmic microtubule network and display severe morphological defects in genetic backgrounds that produce long cells.The tip-specific marker, Tea1p, is mislocalized in both tea2-1 and tea2Delta cells, indicating that Tea2p function is necessary for proper localization of Tea1p.These results suggest that this kinesin promotes microtubule growth, possibly through interactions with the microtubule end, and that it is important for establishing and maintaining polarized growth along the long axis of the cell.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347, USA. browninh@icrf.icnet.uk

ABSTRACT
Cytoplasmic microtubules are critical for establishing and maintaining cell shape and polarity. Our investigations of kinesin-like proteins (klps) and morphological mutants in the fission yeast Schizosaccharomyces pombe have identified a kinesin-like gene, tea2(+), that is required for cells to generate proper polarized growth. Cells deleted for this gene are often bent during exponential growth and initiate growth from improper sites as they exit stationary phase. They have a reduced cytoplasmic microtubule network and display severe morphological defects in genetic backgrounds that produce long cells. The tip-specific marker, Tea1p, is mislocalized in both tea2-1 and tea2Delta cells, indicating that Tea2p function is necessary for proper localization of Tea1p. Tea2p is localized to the tips of the cell and in a punctate pattern within the cell, often coincident with the ends of cytoplasmic microtubules. These results suggest that this kinesin promotes microtubule growth, possibly through interactions with the microtubule end, and that it is important for establishing and maintaining polarized growth along the long axis of the cell.

Show MeSH