HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 275, Issue 27, 20235-20238, July 7, 2000

This Article Abstract Full Text (PDF) All Versions of this Article: 275/27/20235    most recent C000213200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ghiselli, G. Articles by Iozzo, R. V. Search for Related Content PubMed PubMed Citation Articles by Ghiselli, G. Articles by Iozzo, R. V. Social Bookmarking                      What's this?

ACCELERATED PUBLICATION
Overexpression of Bamacan/SMC3 Causes Transformation^*

Giancarlo Ghiselli and Renato V. Iozzo^§¶

From the  Department of Pathology, Anatomy and Cell Biology, and ^§ Program in Cell Biology, Kimmel Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107

Received for publication, March 30, 2000, and in revised form, May 2, 2000

	ABSTRACT

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

Bamacan can occur in certain cell types as either a secreted proteoglycan assembled into basement membranes or as an intracellular protein known as structural maintenance of chromosome 3 (SMC3). To assess the role of this protein in tumorigenesis, we investigated whether induced overexpression of bamacan/SMC3 could transform normal fibroblasts. We generated a full-length cDNA encoding the entire mouse bamacan/SMC3 and demonstrated appropriate transcription and translation into a 146-kDa protein. All the NIH and Balb/c 3T3 murine fibroblasts overexpressing this bamacan/SMC3 transgene generated foci of transformation and acquired anchorage-independent growth. The increased levels of bamacan/SMC3 expression achieved in the transfected fibroblasts were the same as those detected in a series of spontaneously transformed murine and human colon carcinoma cells. Moreover, a 3-4-fold overexpression of bamacan/SMC3 was detected in ~70% of human colon carcinoma specimens from matched pairs (n = 19, p < 0.0002) and in a cohort of intestinal tumors from Apc-deficient Min/+ mice. These results support the concept that deregulated expression of bamacan/SMC3 is involved in cell transformation.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

Originally isolated from the embryonic parietal yolk sac as a high density chondroitin sulfate proteoglycan (1, 2), bamacan was subsequently identified to be a component of the renal mesangial matrix (3), the basement membrane of other tissues (4), and tumor matrix (5). The murine protein is encoded by 31 exons distributed along ~45 kilobase pairs of genomic DNA (6) and is highly conserved across species (7-9). Secondary structure analysis of the protein reveals three globular domains intercalated by two -helix coiled-coils in an antiparallel arrangement folded at a flexible hinge (10). The terminal ends harbor five glycanation sites to which the attachment of glycosaminoglycan side GAG chains is possible, and a P-loop and a DA-box motif that may act cooperatively to bind ATP and DNA, respectively (11). These features and the high sequence homology with the structural maintenance of chromosome 3 (SMC3)¹ protein led us to conclude that bamacan is a member of this family (6). SMC3 proteins are involved in the formation of at least two multimeric complexes within the nucleus. The first, termed "cohesin", is involved in chromosome cohesion and prevents premature sister chromatid separation (12). The second, called RC-1, is involved in DNA repair and has DNA ligase and polymerase activity (13). The formation of a third complex has been postulated on the basis that bamacan/SMC3 can be co-isolated with proteins involved in microtubule dynamics (9). Finally, the protein has a functional leucine zipper domain that binds Mxi and Max, two nuclear proteins that interact with Myc and modulate its transcriptional activity (14).

We previously detected increased bamacan/SMC3 mRNA levels in several transformed cell lines (6), suggesting that the amplified product of the normal gene may play a role in tumorigenesis. To directly test this hypothesis we investigated the effects of transgenic overexpression of bamacan/SMC3 in NIH and Balb/c 3T3 mouse fibroblasts. All the cells overexpressing bamacan/SMC3 showed evidence of transformation, including anchorage-independent growth and foci of transformation. Moreover, all the stably transfected clones exhibited increased levels of bamacan/SMC3 identical to those of spontaneously transformed human colon carcinoma cell lines and colon cancer specimens, as well as intestinal tumors from Apc-deficient Min/+ mice. Thus, deregulated expression of bamacan/SMC3 may be directly linked to the complex process of oncogenesis.

	EXPERIMENTAL PROCEDURES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

Materials and Cells-- [-³²P]dATP (~3000 Ci/mmol) and [³⁵S]methionine (~1000 Ci/mmol) were obtained from Amersham Pharmacia Biotech. The transcription/translation TNT T7 kit was from Promega. Rabbit anti-mouse bamacan/SMC3 antibody was kindly provided by A. Strunnikov. Cell lines include: NIH and Balb/c 3T3 mouse fibroblasts that display contact inhibition and serum dependence; a non-tumorigenic colon epithelial cell line originated from p53 / mice (15); CMT-93 mouse colorectal carcinoma cells; and the human colon carcinoma cell lines HCT116, WiDr, SW480, LOVO, and CaCo2 (American Type Culture Collection). Nineteen human colon carcinoma specimens and matched normal tissues, and five Min/+ animals were also investigated.

Generation of Full-length Bamacan cDNA and Expression Vectors-- Full-length bamacan cDNA was assembled by sequential ligation of EST clones AA144255, AA164123, and AA119169 and of a cDNA fragment generated by reverse transcriptase polymerase chain reaction (RT-PCR). Because of the lack of suitable subcloning vectors, one was generated from pBS-KS+ by replacement of the multiple cloning site between the KpnI and the NotI with a 24-bp oligonucleotide corresponding to the restriction sites KpnI, SpeI, XhoI, NcoI, SalI, NotI (5' to 3'). These new sites allowed the sequential subcloning of the various cDNAs. The coding region of bamacan/SMC3 was inserted at the NotI/EcoRV sites of the pcDNA3.1 myc-His (Invitrogen). A pLXSN retroviral expression vector (CLONTECH) was also constructed. All ligations were verified by restriction map digestion and DNA sequencing.

Stable Transfection, Tumorigenic Assays, and Northern and Western Blotting-- Balb/c 3T3 cells were transfected with 50 µg of expression vector by electroporation (230 V, 1080 microfarads/s). After 2 days, G418 (250 µg/ml) was added and drug selection carried out for 2 weeks. Single colonies were ring-cloned and expanded. Ten days later, RNA was extracted from a first group of cells whereas the parallel group was fixed in 70% ethanol, stained with 0.2% methylene blue, and analyzed for the presence of foci of transformation. Growth in soft agar and colony formation was determined by culturing 3 × 10³ cells between two layers of 0.4% (bottom) and 0.2% (top) agar solution. Colony size was scored at 14 and 21 days by counting the number of cell aggregates >100 µm diameter in several randomly selected 9-mm² areas. Bamacan mRNA levels were assessed by Northern blotting as described previously (6). Cellular levels of bamacan/SMC3 were assessed by Western immunoblotting. Briefly, the cells were solubilized in phosphate-buffered saline containing 0.1% Triton X-100 and a mixture of protease inhibitors Complete^TM (Roche Molecular Biochemicals), separated by SDS-PAGE, and immunoblotted using the primary antibody at 1:300 dilution and the secondary (peroxidase-conjugated anti-rabbit IgG) at 1:17,000 dilution, followed by chemiluminescence (Pierce) detection.

	RESULTS

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

Generation of an Expression Vector Encoding the Full-length Bamacan/SMC3 cDNA and in Vitro Transcription/Translation-- To assess the role of bamacan in tumorigenesis, we first needed to construct a full-length eukaryotic expression vector. The strategy for the assembly of full-length mouse bamacan/SMC3 cDNA, and the pcDNA3.1 and retroviral pLXSN expression vectors, is illustrated in Fig. 1, A and B. The full-length cDNA encompasses 3672 bp and encodes 1206 amino acid residues, including a C-terminal stretch of 11 residues that are not present in the rat sequence (7). To assess whether the predicted open reading frame of the bamacan/SMC3 cDNA could be translated into a protein, the pcDNA3.1 vector was tested in a rabbit reticulocyte lysate transcription-translation system using [³⁵S]methionine as the precursor. The results showed a single protein of the expected size of ~146 kDa (Fig. 1C, lanes 1 and 3). Lack of the expression vector in the reaction mixture generated no detectable protein (Fig. 1C, lane 2), whereas addition of the vector encoding luciferase gave the predicted band of ~60 kDa (Fig. 1C, lane 4). Thus, the full-length cDNA encoding bamacan protein is appropriately transcribed and translated into a protein of size (146 kDa) identical to the native bamacan/SMC3.

View larger version (24K): [in this window] [in a new window]   Fig. 1.   Generation of full-length bamacan expression vectors and in vitro transcription/translation. A, schematic representation of the various EST clones and the RT-PCR generated insert that were sequentially ligated into pBS-KS+. B, expression vectors constructed by ligating bamacan/SMC3 cDNA into a myc-His-tagged pcDNA3.1 or pLXSN retroviral vector. C, autoradiograph of a 7% SDS-PAGE from the in vitro transcription-translation of the bamacan/SMC3 pcDNA3.1 vector using [35S]methionine as the precursor. Notice the presence of a ~146-kDa band in lanes 1 and 3, in the presence or absence or protease inhibitors, respectively. Lane 2 lacks the vector, whereas lane 4 is the positive control vector harboring the luciferase gene encoding a ~60-kDa protein.

Overexpression of Bamacan/SMC3 Causes Transformation of Normal Murine Cells-- Because bamacan mRNA levels are elevated in a number of tumorigenic cell lines (6), we investigated whether overexpression of this protein could directly produce cell transformation. For this purpose, normal NIH and Balb/c 3T3 fibroblasts, which are both contact inhibited, were transfected with the pcDNA3.1 bamacan/SMC3 expression vector. Of the 22 clones obtained, 3 displayed ~4-fold bamacan overexpression (Fig. 2A). Eighteen of these clones were tested for the ability to generate colonies in soft agar. In these experiments, clones 9 and 22, i.e. those overexpressing bamacan mRNA, displayed anchorage-independent growth and generated 31 ± 4 (n = 3) and 23 ± 4 (n = 4) colonies >100 µm diameter after 3 weeks of growth (Fig. 2B). No colonies were observed in the remaining 16 G418-resistant clones expressing normal bamacan mRNA and in wild type cells. Growth kinetics experiments revealed that the overexpressing clones had lost contact inhibition, confirming they had acquired a transformed phenotype. Initial growth rate did not differ significantly between overexpressing and normally expressing bamacan clones. However, upon reaching confluence, normally expressing clones significantly reduced their duplication rate to ~0.2 doubling/day, whereas clones 9 and 22 maintained a vigorous duplication activity (~0.6-1 doubling/day) (Fig. 2C). All the G418-resistant clones were tested in a transformation focus assay. Whereas wild type and G418-resistant clones expressing normal bamacan/SMC3 mRNA levels did not generate foci of transformation, clones 9 and 22 generated numerous foci of transformation (Fig. 2D). Moreover, cells expressing normal bamacan level (clone 6) formed a uniform monolayer, whereas cells overexpressing bamacan exhibited an haphazard arrangement and lacked contact inhibition (Fig. 2E). In agreement with the Northern blotting data, the cellular levels of bamacan/SMC3 protein in the overexpressing clones were ~3-fold higher than control (Fig. 2F). In contrast, no bamacan levels could be detected in the media conditioned by either wild type or transfected cells (not shown).

View larger version (45K): [in this window] [in a new window]   Fig. 2.   Overexpression of bamacan/SMC3 causes transformation of Balb-c 3T3 fibroblasts. A, Northern blotting of total RNA extracted from clone 6 (a G418-resistant but lacking bamacan/SMC3 expression) and two overexpressing clones. For loading comparison, the ethidium bromide-stained ribosomal RNA (rRNA) is shown in the lower panel. B, growth in soft agar as determined after 21 days of growth. C, growth curve of three individual clones as indicated. D, focus assay of clones as in A. The wild type was identical to clone 6 (not shown). E, morphological changes and lack of contact inhibition in cells overexpressing bamacan/SMC3. F, Western immunoblotting of total cell extracts (~20 µg of protein each) of clone 6 (lane 1) and clone 9 (lane 2) using a rabbit antibody raised against Pro1137-Ser1157 region of murine bamacan.

To corroborate the transformation potential of bamacan, we performed additional experiments in which bamacan/SMC3 gene was delivered into NIH 3T3 cells using pLXSN retroviral vector. Among 12 clones isolated, one displayed increased bamacan/SMC3 mRNA (clone 1, Fig. 3A) and protein (Fig. 3B) levels. As in the cells transfected with the pcDNA3.1 vector, the retroviral clones overexpressing bamacan formed numerous foci of transformation (Fig. 3C).

View larger version (44K): [in this window] [in a new window]   Fig. 3.   Overexpression of bamacan/SMC3 causes transformation of NIH 3T3 fibroblasts. A, Northern blotting of five independent G418-resistant clones transfected with pLXSN retroviral bamacan/SMC3 expression vector. For loading comparison, the ethidium bromide-stained ribosomal RNA (rRNA) is shown in the lower panel. B, Western immunoblotting of bamacan/SMC3 levels (20 µg of cellular protein) in clone 1, a bamacan overexpressing clone, and clone 2 as representative of one of 12 clones expressing normal bamacan/SMC3 levels. C, focus assay for clones 1 and 2.

Collectively, these findings support the conclusion that induction of intracellular bamacan/SMC3 expression is sufficient to cause transformation of normal, contact-inhibited fibroblasts.

Bamacan Expression Is Abnormally Elevated in Colon Carcinoma Cell Lines and Colon Carcinoma Tissues-- To assess whether the degree of bamacan/SMC3 expression that caused transformation of 3T3 fibroblast was similar to that of established neoplastic cells, a series of mouse and human neoplastic cell lines was examined. A non-tumorigenic colon epithelial cell line originated from p53 / mice was used as control. These cells do not grow in soft agar and do not generate tumors in either SCID or immunocompetent syngeneic mice (15). Compared with the expression in this primary colon cell line, a 5-fold increase in bamacan/SMC3 expression was observed inCMT-93 mouse colorectal carcinoma cells (Fig. 4A). Similar results were obtained when normal colon tissue and the matching neoplastic tissue from a patient with colon carcinoma were investigated (Fig. 4B). Five independent human colon carcinoma cell lines displayed the same degree of bamacan overexpression as the colon carcinoma sample (Fig. 4B), suggesting that the phenomenon does not simply reflect the high rate of mitosis of the cells held in culture. We then compared bamacan/SMC3 expression in matched neoplastic and normal colon tissues (n = 19). About 70% of the colon carcinomas displayed a significant increase in bamacan/SMC3 steady state mRNA levels (Fig. 4C). In the colon carcinomas, bamacan mRNA levels were 3.7 ± 0.4 (mean ± S.E.) higher than matched control tissues (p < 0.0002 by paired Student's t test) (Fig. 4D).

View larger version (32K): [in this window] [in a new window]   Fig. 4.   Bamacan expression is abnormally elevated in colon carcinoma cell lines, human colon carcinoma, and in intestinal tumors from Min/+ mice. A, Northern blotting of murine normal colon epithelium and colorectal carcinoma CMT-93 cell line. The blot was hybridized with a murine bamacan-specific 32P-labeled cDNA (P144). For loading comparison, the ethidium bromide-stained ribosomal RNA (rRNA) is shown in the lower panel. B, Northern blotting of human colon and colon carcinoma tissues and five independent colon carcinoma cell lines as indicated. The blot was hybridized with a human bamacan-specific 32P-labeled cDNA (P554). C, Northern blotting of representative human colon (N) and colon carcinoma (T) specimens. D, quantification of 19 matched samples from normal and neoplastic colon tissues. Films were scanned with the NIH Scion Image software, which provides a direct numeric output for the densitometric profiles. Data are plotted as arbitrary intensity units corrected for the amount of rRNA. E, Northern blotting of bamacan expression in various tissues from Min/+ mice. The intestine of 8-month-old Min/+ mice were dissected under a microscope and the adenomatous areas carefully separated from normal tissue. Other collected tissues are as indicated. F, abundance of bamacan mRNA in the different specimens calculated as relative to the matching normal small intestine. In E and F, lanes 1, 3, 5, and 7 represent tissues from a C57BL/6J Apc +/+, whereas lanes 2, 4, 6, and 8 represent tissues from a Min/+ mouse. The adenomatous tissue is in lane 2. Samples in lanes 9-12 are from two Min/+ mice and represent normal (lanes 9 and 11) and adenomatous tissue (lanes 10 and 12). G, Western immunoblotting of two Min/+ mice (~20 µg of protein each) using anti-bamacan antiserum. Lanes 1 and 3 represent normal small intestine, whereas lanes 2 and 4 are the matching neoplastic tissues.

Bamacan Is Overexpressed in Intestinal Tumors of APC-deficient Min/+ Mice-- There is convincing evidence that most human colon carcinomas exhibit loss of heterozygosity at the Apc locus coding for the APC tumor suppressor gene (16). In mice, a non-sense mutation (Min allele) of the Apc gene causes the development of multiple intestinal neoplasia (Min/+) (17). As in humans, the adenomas in Min/+ mice show loss of the wild type Apc allele. Therefore, we sought to establish whether bamacan is also overexpressed in adenomas from Min/+ mice. Careful examination of five animals of eight months of age revealed that all had advanced intestinal neoplasia covering ~60% of the small intestine. In agreement with the data presented above, there was a ~4-fold elevation of bamacan/SMC3 expression in the tumors (Fig. 4, E and F). This elevation was specific inasmuch as no difference in bamacan/SMC3 mRNA levels was detected in the proximal or distal colon, or in the testis, when compared with the levels expressed in the same tissues of sex- and age-matched Apc +/+ C57BL/6J mice. The increased expression of bamacan mRNA in the intestinal tumors was confirmed by Western immunoblotting, which showed a ~4-fold increase of bamacan protein in the neoplastic tissues (Fig. 4G). These findings support the concept that an APC-related pathway may mediate bamacan up-regulation in colon tumors.

	DISCUSSION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

The results of this study strengthen the notion that bamacan/SMC3 may play an important role in tumorigenesis (6). The degree of gene activation that leads to cell transformation in normal mouse cells is comparable with that observed in a series of colon carcinomas cell lines and in the majority of human colon carcinoma specimens. The average 4-fold increase supports the idea that bamacan/SMC3 biosynthesis is tightly regulated in normal cells and that an imbalance in the amounts of this protein can directly affect the transformation program.

Several functional roles have been assigned to eukaryotic bamacan/SMC3 for which it is possible to envision mechanisms linking its overexpression to cell growth disregulation. The involvement of this protein in chromatid cohesion during metaphase points to a first mechanism (18). The protein is part of the cohesin multimeric protein complex that prevents the premature separation of sister chromatids (12). This process occurs in an orderly fashion during anaphase under the control of the cell cycle machinery and, if improperly executed, could result in the segregation of both sisters to the same cell thereby generating an aneuploid karyotype (19). Bamacan/SMC3 is also a necessary component, together with SMC1, of a multimeric complex named RC1 that has DNA recombination/renaturation, DNA ligase, and DNA polymerase activities (13). The enzymes act in concert to repair a gapped or deleted DNA. These different functions of bamacan/SMC3 are mediated by its ability to bind palindromic DNA sequences through its C terminus and by allowing the formation of protein-DNA structures that are accessible to the action of DNA-modifying enzymes (20, 21).

The functional significance of proteoglycan core protein in the nucleus and at the same time extracellularly eludes a simple interpretation. Proteoglycans can affect cell growth and differentiation by acting through different signaling pathways (22). Increased levels of proteoglycans with altered composition have been reported in human tumors, primarily those of epithelial origin, such as breast, colon, and lung. Bamacan proteoglycan has been shown to be a normal constituent of the basement membranes in several tissues (3) and is, thus, plausible that its secretion is a physiological event. Whether abnormal accumulation of extracellular bamacan occurs during tumor formation and the exact extracellular function of this protein needs to be established in future studies.

In conclusion, our results support the concept that overexpression of bamacan/SMC3 alone is sufficient to initiate cell transformation in normal fibroblasts. Thus, abnormal expression of bamacan/SMC3 gene may disrupt the formation of the multimeric protein complexes within the nucleus which could directly contribute to the genesis of a transformed phenotype.

	ACKNOWLEDGEMENTS

We thank I. Eichstetter for excellent technical assistance, A. Strunnikov for the gift of anti-bamacan/SMC3 antiserum, and L. Siracusa for providing the Min/+ mice.

	FOOTNOTES

^* This work was supported by National Institutes of Health Grants RO1 CA39481 and RO1 CA47282 (to R. V. I).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: Dept. of Pathology, Anatomy and Cell Biology, Rm. 249, JAH, Thomas Jefferson University, 1020 Locust St., Philadelphia, PA 19107. E-mail: iozzo@lac.jci. tju.edu.

Published, JBC Papers in Press, May 3, 2000, DOI 10.1074/jbc.C000213200

	ABBREVIATIONS

The abbreviations used are: SMC3, structural maintenance of chromosome protein 3; PAGE, polyacrylamide gel electrophoresis; RT-PCR, reverse transcriptase polymerase chain reaction; bp, base pair(s); PAGE, polyacrylamide gel electrophoresis; EST, expressed sequence tag.

	REFERENCES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS DISCUSSION REFERENCES

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				S. K. Olson, J. R. Bishop, J. R. Yates, K. Oegema, and J. D. Esko Identification of novel chondroitin proteoglycans in Caenorhabditis elegans: embryonic cell division depends on CPG-1 and CPG-2 J. Cell Biol., June 19, 2006; 173(6): 985 - 994. [Abstract] [Full Text] [PDF]

				E. M. Assmann, M. R. Alborghetti, M. E. R. Camargo, and J. Kobarg FEZ1 Dimerization and Interaction with Transcription Regulatory Proteins Involves Its Coiled-coil Region J. Biol. Chem., April 14, 2006; 281(15): 9869 - 9881. [Abstract] [Full Text] [PDF]

				M. P. Bard, J. P. Hegmans, A. Hemmes, T. M. Luider, R. Willemsen, L.-A. A. Severijnen, J. P. van Meerbeeck, S. A. Burgers, H. C. Hoogsteden, and B. N. Lambrecht Proteomic Analysis of Exosomes Isolated from Human Malignant Pleural Effusions Am. J. Respir. Cell Mol. Biol., July 1, 2004; 31(1): 114 - 121. [Abstract] [Full Text] [PDF]

				E. Lara-Pezzi, N. Pezzi, I. Prieto, I. Barthelemy, C. Carreiro, A. Martinez, A. Maldonado-Rodriguez, M. Lopez-Cabrera, and J. L. Barbero Evidence of a Transcriptional Co-activator Function of Cohesin STAG/SA/Scc3 J. Biol. Chem., February 20, 2004; 279(8): 6553 - 6559. [Abstract] [Full Text] [PDF]

				J.-F. Lambert, M. Liu, G. A. Colvin, M. Dooner, C. I. McAuliffe, P. S. Becker, B. G. Forget, S. M. Weissman, and P. J. Quesenberry Marrow Stem Cells Shift Gene Expression and Engraftment Phenotype with Cell Cycle Transit J. Exp. Med., June 2, 2003; 197(11): 1563 - 1572. [Abstract] [Full Text] [PDF]

				G. Ghiselli, N. Coffee, C. E. Munnery, R. Koratkar, and L. D. Siracusa The Cohesin SMC3 Is a Target the for {beta}-Catenin/TCF4 Transactivation Pathway J. Biol. Chem., May 23, 2003; 278(22): 20259 - 20267. [Abstract] [Full Text] [PDF]

				D. R. Carson and M. F. Christman Evidence that replication fork components catalyze establishment of cohesion between sister chromatids PNAS, July 17, 2001; 98(15): 8270 - 8275. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) All Versions of this Article: 275/27/20235    most recent C000213200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ghiselli, G. Articles by Iozzo, R. V. Search for Related Content PubMed PubMed Citation Articles by Ghiselli, G. Articles by Iozzo, R. V. Social Bookmarking                      What's this?