Volume 271, Number 45, Issue of November 8, 1996 pp. 28414-28421
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Use of Normal and Transgenic Mice to Examine the Relationship between Terminal Differentiation of Intestinal Epithelial Cells and Accumulation of Their Cell Cycle Regulators

(Received for publication, June 10, 1996)

Chitra Chandrasekaran , Craig M. Coopersmith ^§ and Jeffrey I. Gordon

From the Departments of  Molecular Biology and Pharmacology and ^§ Surgery, Washington University School of Medicine, St. Louis, Missouri 63110

A spatially well organized continuum of proliferation, differentiation, and death is displayed along crypt-villus units in the adult mouse small intestine. This continuum provides an opportunity to examine in vivo the mechanisms by which proliferative status changes as a function of cellular differentiation. Immunohistochemical studies of normal FVB/N mice revealed that as epithelial cells complete their terminal differentiation during a 48-72-h migration up villi, there is a marked and rapid fall in the levels of two important regulators of the G₁/S transition, cyclin D₁ and cyclin-dependent kinase (cdk) 2. However, cellular levels of their partners, cdk4 and cyclin E, remain unchanged as does the level of pRB. Adult FVB/N transgenic mice were studied that contained an intestinal fatty acid binding protein gene promoter (Fabpi) linked to wild type Simian virus 40 large T antigen (SV40 TAg^Wt) or a mutant TAg with Lys for Glu substitutions at residues 107 and 108 (SV40 TAg^K107/8) that fails to bind pRB and related pocket proteins. Both transgenes are expressed only in villus enterocytes. SV40 TAg^Wt causes these terminally differentiated cells to re-enter the cycle. Re-entry is accompanied by a reduction in un/hypophosphorylated pRB, an induction of cyclin D₁ and cdk2, but no change in cdk4, cyclin E, or E2F-1. In contrast, SV40 TAg^K107/8 fails to induce re-entry and does not produce changes in un/hypophosphorylated pRB, cyclin D₁, or cdk2 accumulation. These results suggest that un/hypophosphorylated pRB is an important mediator of the cell cycle arrest that normally occurs as enterocytes exit the crypt and complete their differentiation. Fabpi-directed expression of E2F-1 does not cause villus enterocytes to return to the cell cycle, alter their suppression of cyclin D₁ or cdk2, or affect their state of differentiation, emphasizing the insensitivity of these cells to the effects of E2F-1. Analyses of p53^/ and p53^+/+ mice containing Fabpi-SV40 TAg^Wt and Fabpi-SV40 TAg^K107/8 established that the proliferation induced by SV40 TAg^Wt does not require p53 and is associated with increased (p53-independent) apoptosis. The presence of cyclin E and cdk4 in differentiating villus enterocytes emphasizes that these cells retain part of their proliferative heritage expressed 24-72 h earlier in the crypt. The data suggest that down-regulation of cdk2 and/or cyclin D₁ expression may be important for control of proliferative status and/or execution of terminal differentiation.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. T. Saenz-Robles, J. A. Markovics, J.-L. Chong, R. Opavsky, R. H. Whitehead, G. Leone, and J. M. Pipas Intestinal Hyperplasia Induced by Simian Virus 40 Large Tumor Antigen Requires E2F2 J. Virol., December 1, 2007; 81(23): 13191 - 13199. [Abstract] [Full Text] [PDF]

				A. V. Rathi, M. T. Saenz Robles, and J. M. Pipas Enterocyte Proliferation and Intestinal Hyperplasia Induced by Simian Virus 40 T Antigen Require a Functional J Domain J. Virol., September 1, 2007; 81(17): 9481 - 9489. [Abstract] [Full Text] [PDF]

				A. A. Hizli, A. R. Black, M. A. Pysz, and J. D. Black Protein Kinase C {alpha} Signaling Inhibits Cyclin D1 Translation in Intestinal Epithelial Cells J. Biol. Chem., May 26, 2006; 281(21): 14596 - 14603. [Abstract] [Full Text] [PDF]

				M. Plateroti, E. Kress, J. I. Mori, and J. Samarut Thyroid Hormone Receptor {alpha}1 Directly Controls Transcription of the {beta}-Catenin Gene in Intestinal Epithelial Cells Mol. Cell. Biol., April 15, 2006; 26(8): 3204 - 3214. [Abstract] [Full Text] [PDF]

				K. Haigis, J. Sage, J. Glickman, S. Shafer, and T. Jacks The Related Retinoblastoma (pRb) and p130 Proteins Cooperate to Regulate Homeostasis in the Intestinal Epithelium J. Biol. Chem., January 6, 2006; 281(1): 638 - 647. [Abstract] [Full Text] [PDF]

				S. Kim, A. J. Schein, and J. A. Nadel E-cadherin promotes EGFR-mediated cell differentiation and MUC5AC mucin expression in cultured human airway epithelial cells Am J Physiol Lung Cell Mol Physiol, December 1, 2005; 289(6): L1049 - L1060. [Abstract] [Full Text] [PDF]

				J. A. Markovics, P. A. Carroll, M. T. S. Robles, H. Pope, C. M. Coopersmith, and J. M. Pipas Intestinal Dysplasia Induced by Simian Virus 40 T Antigen Is Independent of p53 J. Virol., June 15, 2005; 79(12): 7492 - 7502. [Abstract] [Full Text] [PDF]

				J. R. Gum Jr., J. W. Hicks, S. C. Crawley, S. C. Yang, A. D. Borowsky, C. M. Dahl, S. Kakar, D. H. Kim, R. D. Cardiff, and Y. S. Kim Mice Expressing SV40 T Antigen Directed by the Intestinal Trefoil Factor Promoter Develop Tumors Resembling Human Small Cell Carcinoma of the Colon Mol. Cancer Res., September 1, 2004; 2(9): 504 - 513. [Abstract] [Full Text] [PDF]

				C. Ratineau, C. Bernard, G. Poncet, M. Blanc, C. Josso, S. Fontaniere, A. Calender, J. A. Chayvialle, C.-X. Zhang, and C. Roche Reduction of Menin Expression Enhances Cell Proliferation and Is Tumorigenic in Intestinal Epithelial Cells J. Biol. Chem., June 4, 2004; 279(23): 24477 - 24484. [Abstract] [Full Text] [PDF]

				H. Yang, P. A. Antony, B. E. Wildhaber, and D. H. Teitelbaum Intestinal Intraepithelial Lymphocyte {gamma}{delta}-T Cell-Derived Keratinocyte Growth Factor Modulates Epithelial Growth in the Mouse J. Immunol., April 1, 2004; 172(7): 4151 - 4158. [Abstract] [Full Text] [PDF]

				I. U. Mysorekar, R. G. Lorenz, and J. I. Gordon A Gnotobiotic Transgenic Mouse Model for Studying Interactions between Small Intestinal Enterocytes and Intraepithelial Lymphocytes J. Biol. Chem., September 27, 2002; 277(40): 37811 - 37819. [Abstract] [Full Text] [PDF]

				C. Ratineau, M. W. Petry, H. Mutoh, and A. B. Leiter Cyclin D1 Represses the Basic Helix-Loop-Helix Transcription Factor, BETA2/NeuroD J. Biol. Chem., March 8, 2002; 277(11): 8847 - 8853. [Abstract] [Full Text] [PDF]

				James. R. Gum Jr., James. W. Hicks, A. M. Gillespie, J. L. Rius, P. A. Treseler, S. C. Kogan, E. J. Carlson, C. J. Epstein, and Y. S. Kim Mouse Intestinal Goblet Cells Expressing SV40 T Antigen Directed by the MUC2 Mucin Gene Promoter Undergo Apoptosis upon Migration to the Villi Cancer Res., April 1, 2001; 61(8): 3472 - 3479. [Abstract] [Full Text]

				M. R. Frey, J. A. Clark, O. Leontieva, J. M. Uronis, A. R. Black, and J. D. Black Protein Kinase C Signaling Mediates a Program of Cell Cycle Withdrawal in the Intestinal Epithelium J. Cell Biol., November 13, 2000; 151(4): 763 - 778. [Abstract] [Full Text] [PDF]

				A. J. M. Watson and D. M. Pritchard Lessons From Genetically Engineered Animal Models. VII. Apoptosis in intestinal epithelium: lessons from transgenic and knockout mice Am J Physiol Gastrointest Liver Physiol, January 1, 2000; 278(1): G1 - G5. [Abstract] [Full Text] [PDF]

				Q. Ding, Q. Wang, Z. Dong, and B. M. Evers Characterization and regulation of E2F activity during Caco-2 cell differentiation Am J Physiol Cell Physiol, January 1, 2000; 278(1): C110 - C117. [Abstract] [Full Text] [PDF]

				C. P. Sparrow, S. Patel, J. Baffic, Y.-S. Chao, M. Hernandez, M.-H. Lam, J. Montenegro, S. D. Wright, and P. A. Detmers A fluorescent cholesterol analog traces cholesterol absorption in hamsters and is esterified in vivo and in vitro J. Lipid Res., October 1, 1999; 40(10): 1747 - 1757. [Abstract] [Full Text]

				R. J. Bova, D. I. Quinn, J. S. Nankervis, I. E. Cole, B. F. Sheridan, M. J. Jensen, G. J. Morgan, C. J. Hughes, and R. L. Sutherland Cyclin D1 and p16INK4A Expression Predict Reduced Survival in Carcinoma of the Anterior Tongue Clin. Cancer Res., October 1, 1999; 5(10): 2810 - 2819. [Abstract] [Full Text] [PDF]

				G Svineng and S Johansson Integrin subunits (beta)1C-1 and (beta)1C-2 expressed in GD25T cells are retained and degraded intracellularly rather than localised to the cell surface J. Cell Sci., January 12, 1999; 112(24): 4751 - 4761. [Abstract] [PDF]

				J. M. Carethers Cell checkpoints and enterocyte differentiation: a recipe for sequential stages Focus on "Caco-2 intestinal cell differentiation is associated with G1 arrest and suppression of CDK2 and CDK4" Am J Physiol Cell Physiol, November 1, 1998; 275(5): C1191 - C1192. [Full Text] [PDF]

				Q.-M. Ding, T. C. Ko, and B. M. Evers Caco-2 intestinal cell differentiation is associated with G1 arrest and suppression of CDK2 and CDK4 Am J Physiol Cell Physiol, November 1, 1998; 275(5): C1193 - C1200. [Abstract] [Full Text] [PDF]

				M. H. Wong, B. Rubinfeld, and J. I. Gordon Effects of Forced Expression of an NH2-terminal Truncated beta -Catenin on Mouse Intestinal Epithelial Homeostasis J. Cell Biol., May 4, 1998; 141(3): 765 - 777. [Abstract] [Full Text] [PDF]

				E. M. Garabedian, L. J. J. Roberts, M. S. McNevin, and J. I. Gordon Examining the Role of Paneth Cells in the Small Intestine by Lineage Ablation in Transgenic Mice J. Biol. Chem., September 19, 1997; 272(38): 23729 - 23740. [Abstract] [Full Text] [PDF]

				C. M. Coopersmith, C. Chandrasekaran, M. S. McNevin, and J. I. Gordon Bi-transgenic Mice Reveal that K-rasVal12 Augments a p53-independent Apoptosis When Small Intestinal Villus Enterocytes Reenter the Cell Cycle J. Cell Biol., July 14, 1997; 138(1): 167 - 179. [Abstract] [Full Text] [PDF]