Correction: Katanin Localization Requires Triplet Microtubules in <i>Chlamydomonas reinhardtii</i>

<p>Correction: Katanin Localization Requires Triplet Microtubules in <i>Chlamydomonas reinhardtii</i></p

The <i>bld2-5</i> and <i>bld2-6</i> strains misplace the cleavage furrow.

<p>A. The ratio of the areas of wild-type sister cells is approximately equal to one (black bars), whereas the ratio of the areas of <i>bld2-5</i> (gray bars) and <i>bld2-6</i> (white bars) sister cells is equal to or greater than one, which suggests a defect in proper placement of the cleavage furrow <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053940#pone.0053940-Preble1" target="_blank">[34]</a>. These results are statistically significant compared by a permutation test <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053940#pone.0053940-Good1" target="_blank">[37]</a>.</p

Katanin localizes to the spindle poles in mitotic cells.

<p>Synchronized cultures were fixed and stained for α-tubulin (red, left panel), HA (green, middle panel), and DNA (blue). Cell cycle stages were determined based on the DNA and tubulin staining patterns. The prometaphase, metaphase, and top row of anaphase are deconvoluted maximum projections of the z-stack. The bottom three rows for anaphase are maximum projections of the z-stack without deconvolution. Scale bar equals 5 µm.</p

Mature basal bodies in the <i>bld2-5</i> strain contain defects in microtubule blade organization.

<p>Selected tomographic slices show the progression from the proximal (A) to the distal end (F) of the basal body. (A) Probasal bodies contain a ring of amorphous material at their proximal base. (B–E; arrowheads) Mature basal bodies (BB1) contain amorphous material that extends distally rather than in a proximal ring. (D–F) The assembly of microtubule blades is also incomplete with singlet, doublet and sometimes triplet microtubules present. (C, D; arrows) The cartwheel structure is observed distally. BB1, mature basal body 1; BB2, mature basal body 2; rMT, rootlet microtubules. Scale bar equals 100 nm and Panels A–F are at the same magnification. Schematic representation of the structure of a wild-type basal body and the defects in <i>bld2-5</i> basal bodies as one moves from the proximal to distal region of the basal body.</p

Numbers of flagella in <i>bld2-5</i>, <i>bld2-6</i> and intragenic revertant strains.

a<p><i>BLD2TG</i> indicates the ε-tubulin transgene described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053940#pone.0053940-Dutcher3" target="_blank">[24]</a>.</p

Basal body mutant strains show supersensitivity to Taxol.

<p>(A) Serial dilution of mutant, rescued, and intragenic revertant strains on control medium and (B) 8 µM Taxol-containing medium. Phase images of cells on media with different Taxol concentrations. (C, G) Wild-type, (D, H) <i>pf15-1</i>, (E, I) <i>bld2-6</i> and (F, J) <i>bld2-6, pf15-1</i> double mutant on 10 µM (C–F) or 6 µM Taxol (G–J) containing medium. The <i>bld2-6, pf15-1</i> double mutant is unable to grow on 6 µM Taxol containing medium compared to the single mutant strains. Scale bar in Panel C equals 10 µm. Panels C–J are at the same magnification.</p

IFT74 localization is aberrant in the <i>bld2</i> alleles.

<p>(A–C) IFT74 and katanin p80 do not colocalize. (D–F) IFT74 (green) localizes to the base of the flagella in wild-type cell as a band as well as to the proximal region of the flagella and partially colocalizes with centrin (red) in the striated fiber at the distal end of the basal body but not along the nucleo basal body connectors at the proximal end (red). (G–I) <i>bld2-5</i> cells show staining at the base of the flagella, however the localization appears reduced compared to wild-type cells. (J–L) IFT74 localizes throughout the cytoplasm in <i>bld2-6</i> cells. In about one-half of the cells examined, IFT74 localizes near the aberrant centrin staining at the nucleus. DNA (blue) is stained with DAPI. Scale bar in Panel A equals 5 µm. Panels A–L are at the same magnification.</p

Centrin localization varies in the <i>bld2-5</i> and <i>bld2-6</i> strains.

<p>(A, B) Wild-type cells with an extended pattern of centrin. (C) Approximately 41% of <i>bld2-5</i> cells have centrin that collapses on the nucleus (n = 70). (D) <i>bld2-5</i> cells with a wild-type centrin pattern. (E, F) The rescued transformant, <i>bld2-5</i>; <i>BLD2</i>, and (G, H) the pseudorevertants, <i>bld2-7</i> and (I, J) <i>bld2-9</i> have extended centrin. (K, L) All <i>bld2-6 cells</i> show centrin collapses on or around the nucleus. Scale bar in panel L equals 10 µm. Panels A–L are at the same magnification.</p

Rootlet microtubules are disorganized in the <i>bld2-5</i> and <i>bld2-6</i> strains.

<p>(A, B) Rootlet microtubules in wild-type cells form a cruicate pattern. (C, D) <i>bld2-5</i> cells show an aberrant number and placement of rootlet microtubules. (E, F) The <i>bld2-5</i>; <i>BLD2</i> strain shows a wild-type rootlet microtubule phenotype (N = 15). Pseudorevertants <i>bld2-7</i> (G, H) and <i>bld2-9</i> (I, J) have a nearly wild-type rootlet microtubule phenotype but splaying occurs at the ends of the microtubules (arrow). (K, L) The <i>tub2-1</i> strain has increased acetylated α-tubulin staining. (M, N) The <i>bld2-6</i> cells have a severe disorganization of rootlet microtubules. Scale bar in Panel A equals 5 µm. Panels A–N are at the same magnification.</p