Abstract
Recently, we have shown that a cancer causing truncation in adenomatous polyposis coli (APC) (APC1-1450) dominantly interferes with mitotic spindle function, suggesting APC regulates microtubule dynamics during mitosis. Here, we examine the possibility that APC mutants interfere with the function of EB1, a plus-end microtubule-binding protein that interacts with APC and is required for normal microtubule dynamics. We show that siRNA-mediated inhibition of APC, EB1, or APC and EB1 together give rise to similar defects in mitotic spindles and chromosome alignment without arresting cells in mitosis; in contrast inhibition of CLIP170 or LIS1 cause distinct spindle defects and mitotic arrest. We show that APC1-1450 acts as a dominant negative by forming a hetero-oligomer with the full-length APC and preventing it from interacting with EB1, which is consistent with a functional relationship between APC and EB1. Live-imaging of mitotic cells expressing EB1-GFP demonstrates that APC1-1450 compromises the dynamics of EB1-comets, increasing the frequency of EB1-GFP pausing. Together these data provide novel insight into how APC may regulate mitotic spindle function and how errors in chromosome segregation are tolerated in tumor cells.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 4609-4622 |
| Number of pages | 14 |
| Journal | Molecular Biology of the Cell |
| Volume | 16 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 2005 |
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ASJC Scopus subject areas
- Molecular Biology
- Genetics
- Cell Biology
Cite this
APC and EB1 function together in mitosis to regulate spindle dynamics and chromosome alignment. / Green, Rebecca A.; Wollman, Roy; Kaplan, Kenneth B.
In: Molecular Biology of the Cell, Vol. 16, No. 10, 10.2005, p. 4609-4622.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - APC and EB1 function together in mitosis to regulate spindle dynamics and chromosome alignment
AU - Green, Rebecca A.
AU - Wollman, Roy
AU - Kaplan, Kenneth B.
PY - 2005/10
Y1 - 2005/10
N2 - Recently, we have shown that a cancer causing truncation in adenomatous polyposis coli (APC) (APC1-1450) dominantly interferes with mitotic spindle function, suggesting APC regulates microtubule dynamics during mitosis. Here, we examine the possibility that APC mutants interfere with the function of EB1, a plus-end microtubule-binding protein that interacts with APC and is required for normal microtubule dynamics. We show that siRNA-mediated inhibition of APC, EB1, or APC and EB1 together give rise to similar defects in mitotic spindles and chromosome alignment without arresting cells in mitosis; in contrast inhibition of CLIP170 or LIS1 cause distinct spindle defects and mitotic arrest. We show that APC1-1450 acts as a dominant negative by forming a hetero-oligomer with the full-length APC and preventing it from interacting with EB1, which is consistent with a functional relationship between APC and EB1. Live-imaging of mitotic cells expressing EB1-GFP demonstrates that APC1-1450 compromises the dynamics of EB1-comets, increasing the frequency of EB1-GFP pausing. Together these data provide novel insight into how APC may regulate mitotic spindle function and how errors in chromosome segregation are tolerated in tumor cells.
AB - Recently, we have shown that a cancer causing truncation in adenomatous polyposis coli (APC) (APC1-1450) dominantly interferes with mitotic spindle function, suggesting APC regulates microtubule dynamics during mitosis. Here, we examine the possibility that APC mutants interfere with the function of EB1, a plus-end microtubule-binding protein that interacts with APC and is required for normal microtubule dynamics. We show that siRNA-mediated inhibition of APC, EB1, or APC and EB1 together give rise to similar defects in mitotic spindles and chromosome alignment without arresting cells in mitosis; in contrast inhibition of CLIP170 or LIS1 cause distinct spindle defects and mitotic arrest. We show that APC1-1450 acts as a dominant negative by forming a hetero-oligomer with the full-length APC and preventing it from interacting with EB1, which is consistent with a functional relationship between APC and EB1. Live-imaging of mitotic cells expressing EB1-GFP demonstrates that APC1-1450 compromises the dynamics of EB1-comets, increasing the frequency of EB1-GFP pausing. Together these data provide novel insight into how APC may regulate mitotic spindle function and how errors in chromosome segregation are tolerated in tumor cells.
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U2 - 10.1091/mbc.E05-03-0259
DO - 10.1091/mbc.E05-03-0259
M3 - Article
C2 - 16030254
AN - SCOPUS:26244463198
VL - 16
SP - 4609
EP - 4622
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 10
ER -