Adenomatous polyposis coli truncation alters cytoskeletal structure and microtubule stability in early intestinal tumorigenesis.
Partial loss of function of adenomatous polyposis coli (APC) protein by truncation of its carboxy (C)-terminus is an early factor in the development of many sporadic colorectal cancers. In the C57BL/6J Min/+ (Min/+) mouse, an animal with a germline mutation of Apc, we found that APC truncation was associated with reduced enterocyte migration and loss of association and membrane expression of adherens junction proteins. We hypothesized that these defects were related to changes in cytoskeletal function resulting from truncation of the APC C-terminus, which contains microtubule binding regions, as well as putative sites for indirect actin binding. We investigated this further by determining whether APC truncation produced in vivo changes in actin cytoskeletal structure and microtubule stability. The actin cytoskeleton of histologically normal enterocytes from Min/+ mice was compared to that of Apc+/+ (wild-type) mice by confocal indirect immunofluorescence microscopy. We found a significant loss of actin localization at the apical plasma membrane in Min/+ enterocytes. In addition, immunoblotting revealed increased levels of both unstable Tyr-tubulin and alpha-tubulin turnover in Min/+ enterocytes, indicating an alteration in microtubule dynamics. These studies suggest that loss of actin localization and changes in microtubule dynamics may be dominant negative effects of truncated APC. These changes are consistent with the defects in enterocyte migration and junctional complex formation observed in the Min/+ model of early APC-associated colorectal tumorigenesis.
Hughes
Sara A
SA
Department of Surgery, Brigham and Women's Hospital, 75 Francis Street, Boston, MA 02115, USA.
Carothers
Adelaide M
AM
Hunt
Daniel H
DH
Moran
Amy E
AE
Mueller
James D
JD
Bertagnolli
Monica M
MM
D011125:Adenomatous Polyposis Coli / Q000235:genetics*; D025601:Adenomatous Polyposis Coli Protein / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D020895:Enterocytes / Q000166:cytology / Q000502:physiology; D005260:Female; D017491:Genes, APC; D016147:Genes, Tumor Suppressor; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D015739:Nocodazole / Q000494:pharmacology; D010641:Phenotype; D011485:Protein Binding / Q000235:genetics; D012680:Sensitivity and Specificity; D014404:Tubulin / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D025601:Adenomatous Polyposis Coli Protein; D014404:Tubulin; D015739:Nocodazole