Abstract
The colonic crypt is the functional unit of the colon mucosa with a central role in ion and water reabsorption. Under steady-state conditions, the distal colonic crypt harbors a single stem cell at its base that gives rise to highly proliferative progenitor cells that differentiate into columnar, goblet, and endocrine cells. The role of c-Myb in crypt homeostasis has not been elucidated. Here we have studied three genetically distinct hypomorphic c-myb mutant mouse strains, all of which show reduced colonic crypt size. The mutations target the key domains of the transcription factor: the DNA binding, transactivation, and negative regulatory domains. In vivo proliferation and cell cycle marker studies suggest that these mice have a progenitor cell proliferation defect mediated in part by reduced Cyclin E1 expression. To independently assess the extent to which c-myb is required for colonic crypt homeostasis we also generated a novel tissue-specific mouse model to allow the deletion of c-myb in adult colon, and using these mice we show that c-Myb is required for crypt integrity, normal differentiation, and steady-state proliferation.
MeSH Terms
Animals
Cell Cycle
Cell Differentiation
Cell Proliferation
Colon/metabolism
Cyclin E/metabolism
Mice
Mice, Inbred C57BL
Oncogene Proteins/metabolism
Protein Structure, Tertiary
Proto-Oncogene Proteins c-myb/metabolism,physiology
Stem Cells/cytology,metabolism
Transcription Factors/metabolism
Transcriptional Activation
Chemicals
CCNE1 protein, human
Cyclin E
Oncogene Proteins
Proto-Oncogene Proteins c-myb
Transcription Factors
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Malaterre Jordane
Peter MacCallum Cancer Centre and Pathology Department, University of Melbourne, Melbourne VIC 8006, Australia.
Carpinelli Marina
Ernst Matthias
Alexander Warren
Cooke Michael
Sutton Susan
Dworkin Sebastian
Heath Joan K
Frampton Jon
McArthur Grant
Clevers Hans
Hilton Douglas
Mantamadiotis Theo
Ramsay Robert G
References (29)
29 references, click to expand
-
A method for the isolation and culture of human colonic crypts in collagen gels.
In Vitro Cell Dev Biol. 1987 Jun;23(6):436-42
PMID: 3597283
-
Aberrant expression of an amplified c-myb oncogene in two cell lines from a colon carcinoma.
Proc Natl Acad Sci U S A. 1984 Jul;81(14):4534-8
PMID: 6589609
-
Transcriptional activation of the c-myc gene by the c-myb and B-myb gene products.
Oncogene. 1992 Jun;7(6):1233-40
PMID: 1594249
-
Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation.
J Cell Biol. 1992 Nov;119(3):493-501
PMID: 1400587
-
Differentiation and self-renewal in the mouse gastrointestinal epithelium.
Curr Opin Cell Biol. 1994 Dec;6(6):795-803
PMID: 7880525
-
Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control.
Cell. 1995 Aug 25;82(4):675-84
PMID: 7664346
-
A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(Kip1)-deficient mice.
Cell. 1996 May 31;85(5):733-44
PMID: 8646781
-
Colonic expression of c-myb is initiated in utero and continues throughout adult life.
Cell Growth Differ. 1996 Jul;7(7):961-7
PMID: 8809414
-
p21 (WAF1/CIP1) expression is induced in newly nondividing cells in diverse epithelia and during differentiation of the Caco-2 intestinal cell line.
Exp Cell Res. 1996 Sep 15;227(2):171-81
PMID: 8831553
-
Stem cells in gastrointestinal epithelium: numbers, characteristics and death.
Philos Trans R Soc Lond B Biol Sci. 1998 Jun 29;353(1370):821-30
PMID: 9684279
-
c-Myb down-regulation is associated with human colon cell differentiation, apoptosis, and decreased Bcl-2 expression.
Cancer Res. 1998 Nov 15;58(22):5168-75
PMID: 9823328
-
c-Myb is essential for early T cell development.
Genes Dev. 1999 May 1;13(9):1073-8
PMID: 10323859
-
Tumor suppression by p27Kip1 and p21Cip1 during chemically induced skin carcinogenesis.
Oncogene. 1999 Aug 19;18(33):4689-98
PMID: 10467416
-
c-Myb is critical for murine colon development.
Oncogene. 1999 Oct 14;18(42):5821-30
PMID: 10523863
-
Expression of the c-myb proto-oncogene during cellular proliferation.
Nature. 1986 Jan 30-Feb 5;319(6052):374-80
PMID: 3511387
-
Colon epithelial cell differentiation is inhibited by constitutive c-myb expression or mutant APC plus activated RAS.
DNA Cell Biol. 2005 Jan;24(1):21-9
PMID: 15684716
-
c-Myb and p300 regulate hematopoietic stem cell proliferation and differentiation.
Dev Cell. 2005 Feb;8(2):153-66
PMID: 15691758
-
c-Myc is required for the formation of intestinal crypts but dispensable for homeostasis of the adult intestinal epithelium.
Mol Cell Biol. 2005 Sep;25(17):7868-78
PMID: 16107730
-
c-Myb a stem-progenitor cell regulator in multiple tissue compartments.
Growth Factors. 2005 Dec;23(4):253-61
PMID: 16338788
-
Mutations in the MYB intron I regulatory sequence increase transcription in colon cancers.
Genes Chromosomes Cancer. 2006 Dec;45(12):1143-54
PMID: 16977606
-
p27 expression in inflammatory bowel disease-associated neoplasia. Further evidence of a unique molecular pathogenesis.
Am J Pathol. 1999 Nov;155(5):1511-8
PMID: 10550307
-
Analysis of the regulation of the A33 antigen gene reveals intestine-specific mechanisms of gene expression.
J Biol Chem. 2002 Sep 13;277(37):34531-9
PMID: 12114523
-
Targeting c-Myb expression in human disease.
Expert Opin Ther Targets. 2003 Apr;7(2):235-48
PMID: 12667100
-
Cyclin E ablation in the mouse.
Cell. 2003 Aug 22;114(4):431-43
PMID: 12941272
-
Progression through key stages of haemopoiesis is dependent on distinct threshold levels of c-Myb.
EMBO J. 2003 Sep 1;22(17):4478-88
PMID: 12941699
-
B lymphocyte-specific c-Myc expression stimulates early and functional expansion of the vasculature and lymphatics during lymphomagenesis.
Am J Pathol. 2003 Dec;163(6):2233-45
PMID: 14633598
-
Suppressor screen in Mpl-/- mice: c-Myb mutation causes supraphysiological production of platelets in the absence of thrombopoietin signaling.
Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6553-8
PMID: 15071178
-
c-myb Heterozygous mice are hypersensitive to 5-fluorouracil and ionizing radiation.
Mol Cancer Res. 2004 Jun;2(6):354-61
PMID: 15235111
-
A functional c-myb gene is required for normal murine fetal hepatic hematopoiesis.
Cell. 1991 May 17;65(4):677-89
PMID: 1709592