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TACC3 is a microtubule plus end-tracking protein that promotes axon elongation and also regulates microtubule plus end dynamics in multiple embryonic cell types.

Nwagbara BU, Faris AE, Bearce EA, Erdogan B, Ebbert PT, Evans MF, Rutherford EL, Enzenbacher TB, Lowery LA - Mol. Biol. Cell (2014)

Bottom Line: Using high-resolution live-imaging data on tagged +TIPs, we show that TACC3 localizes to the extreme microtubule plus end, where it lies distal to the microtubule polymerization marker EB1 and directly overlaps with the microtubule polymerase XMAP215.TACC3 also plays a role in regulating XMAP215 stability and localizing XMAP215 to microtubule plus ends.Taken together, our results implicate TACC3 as a +TIP that functions with XMAP215 to regulate microtubule plus end dynamics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Boston College, Chestnut Hill, MA 02467.

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TACC3 can act as a +TIP in neuronal growth cones and neural crest cells. (A–C) Expression of mKate2-tubulin (A), GFP-TACC3 (B), and merge (C) in living growth cone (gc). See Figure 3 Supplemental Movie 1. (A′–C′) Magnified time-lapse montages of the boxed regions in A–C shows that GFP-TACC3 localizes to growing MT plus ends. (D) Fluorescence intensity profile of line-scan average from 30 MT plus ends in growth cones. (E) Histogram depicting the distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs in growth cones. (F) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. (G–I) Expression of mKate2-tubulin (G), GFP-TACC3 (H), and merge (I) in living neural crest cell (ncc). See Figure 3 Supplemental Movie 2. (G′–I′) Magnified views of the boxed regions in G–I. See Figure 3 Supplemental Movie 3. (G′′–I′′) Magnified time-lapse montages of the boxed regions in G′–I′. (J) Fluorescence intensity profile of line-scan average from 45 MT plus ends in neural crest cells. (K) Distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs. (L) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. Bar, 1 μm.
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Figure 3: TACC3 can act as a +TIP in neuronal growth cones and neural crest cells. (A–C) Expression of mKate2-tubulin (A), GFP-TACC3 (B), and merge (C) in living growth cone (gc). See Figure 3 Supplemental Movie 1. (A′–C′) Magnified time-lapse montages of the boxed regions in A–C shows that GFP-TACC3 localizes to growing MT plus ends. (D) Fluorescence intensity profile of line-scan average from 30 MT plus ends in growth cones. (E) Histogram depicting the distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs in growth cones. (F) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. (G–I) Expression of mKate2-tubulin (G), GFP-TACC3 (H), and merge (I) in living neural crest cell (ncc). See Figure 3 Supplemental Movie 2. (G′–I′) Magnified views of the boxed regions in G–I. See Figure 3 Supplemental Movie 3. (G′′–I′′) Magnified time-lapse montages of the boxed regions in G′–I′. (J) Fluorescence intensity profile of line-scan average from 45 MT plus ends in neural crest cells. (K) Distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs. (L) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. Bar, 1 μm.

Mentions: To gain further insight into how TACC3 mechanistically affects MT plus end behaviors, we examined the subcellular dynamics of TACC3 within living embryonic cells. Thus we tagged full-length TACC3 with GFP and examined its localization within both growth cones and neural crest cells (Figure 3 and Figure 3 Supplemental Movies 1–3). GFP-tagged TACC3 strongly localized to the growing plus ends of MTs in both cell types (Figure 3, A–C and G–I). In growth cones, the mean length of the GFP-TACC3 plus end accumulation was ∼0.70 μm (Figure 3, D and E; data measured from 70 MTs). GFP-TACC3 was primarily detected on MT plus ends that either appeared to be advancing forward or paused, with 100% of growing MTs displaying detectable GFP-TACC3 plus end localization (Figure 3F). Seventy-seven percent of paused MTs still showed GFP-TACC3 localization, whereas 25% of shrinking MTs also had observable GFP-TACC3 localization. These observations were similar in neural crest cells, although the GFP-TACC3 comets were shorter, with a mean length of 0.53 μm (Figure 3, J and K; data measured from 64 MTs). GFP-TACC3 accumulation was detectable on 100% of growing MTs, 69% of paused MTs, and 25% of shrinking MTs (Figure 3L). Thus we find that GFP-TACC3 robustly tracks plus ends of MTs in vertebrate growth cones and neural crest cells.


TACC3 is a microtubule plus end-tracking protein that promotes axon elongation and also regulates microtubule plus end dynamics in multiple embryonic cell types.

Nwagbara BU, Faris AE, Bearce EA, Erdogan B, Ebbert PT, Evans MF, Rutherford EL, Enzenbacher TB, Lowery LA - Mol. Biol. Cell (2014)

TACC3 can act as a +TIP in neuronal growth cones and neural crest cells. (A–C) Expression of mKate2-tubulin (A), GFP-TACC3 (B), and merge (C) in living growth cone (gc). See Figure 3 Supplemental Movie 1. (A′–C′) Magnified time-lapse montages of the boxed regions in A–C shows that GFP-TACC3 localizes to growing MT plus ends. (D) Fluorescence intensity profile of line-scan average from 30 MT plus ends in growth cones. (E) Histogram depicting the distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs in growth cones. (F) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. (G–I) Expression of mKate2-tubulin (G), GFP-TACC3 (H), and merge (I) in living neural crest cell (ncc). See Figure 3 Supplemental Movie 2. (G′–I′) Magnified views of the boxed regions in G–I. See Figure 3 Supplemental Movie 3. (G′′–I′′) Magnified time-lapse montages of the boxed regions in G′–I′. (J) Fluorescence intensity profile of line-scan average from 45 MT plus ends in neural crest cells. (K) Distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs. (L) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. Bar, 1 μm.
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Figure 3: TACC3 can act as a +TIP in neuronal growth cones and neural crest cells. (A–C) Expression of mKate2-tubulin (A), GFP-TACC3 (B), and merge (C) in living growth cone (gc). See Figure 3 Supplemental Movie 1. (A′–C′) Magnified time-lapse montages of the boxed regions in A–C shows that GFP-TACC3 localizes to growing MT plus ends. (D) Fluorescence intensity profile of line-scan average from 30 MT plus ends in growth cones. (E) Histogram depicting the distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs in growth cones. (F) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. (G–I) Expression of mKate2-tubulin (G), GFP-TACC3 (H), and merge (I) in living neural crest cell (ncc). See Figure 3 Supplemental Movie 2. (G′–I′) Magnified views of the boxed regions in G–I. See Figure 3 Supplemental Movie 3. (G′′–I′′) Magnified time-lapse montages of the boxed regions in G′–I′. (J) Fluorescence intensity profile of line-scan average from 45 MT plus ends in neural crest cells. (K) Distribution of lengths of detectable GFP-TACC3 localization on the plus ends of MTs. (L) Percentage of MT plus ends with GFP-TACC3 localization for different MT dynamics instability states. Bar, 1 μm.
Mentions: To gain further insight into how TACC3 mechanistically affects MT plus end behaviors, we examined the subcellular dynamics of TACC3 within living embryonic cells. Thus we tagged full-length TACC3 with GFP and examined its localization within both growth cones and neural crest cells (Figure 3 and Figure 3 Supplemental Movies 1–3). GFP-tagged TACC3 strongly localized to the growing plus ends of MTs in both cell types (Figure 3, A–C and G–I). In growth cones, the mean length of the GFP-TACC3 plus end accumulation was ∼0.70 μm (Figure 3, D and E; data measured from 70 MTs). GFP-TACC3 was primarily detected on MT plus ends that either appeared to be advancing forward or paused, with 100% of growing MTs displaying detectable GFP-TACC3 plus end localization (Figure 3F). Seventy-seven percent of paused MTs still showed GFP-TACC3 localization, whereas 25% of shrinking MTs also had observable GFP-TACC3 localization. These observations were similar in neural crest cells, although the GFP-TACC3 comets were shorter, with a mean length of 0.53 μm (Figure 3, J and K; data measured from 64 MTs). GFP-TACC3 accumulation was detectable on 100% of growing MTs, 69% of paused MTs, and 25% of shrinking MTs (Figure 3L). Thus we find that GFP-TACC3 robustly tracks plus ends of MTs in vertebrate growth cones and neural crest cells.

Bottom Line: Using high-resolution live-imaging data on tagged +TIPs, we show that TACC3 localizes to the extreme microtubule plus end, where it lies distal to the microtubule polymerization marker EB1 and directly overlaps with the microtubule polymerase XMAP215.TACC3 also plays a role in regulating XMAP215 stability and localizing XMAP215 to microtubule plus ends.Taken together, our results implicate TACC3 as a +TIP that functions with XMAP215 to regulate microtubule plus end dynamics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Boston College, Chestnut Hill, MA 02467.

Show MeSH
Related in: MedlinePlus