Dissection of the asynchronous transport of intestinal microvillar hydrolases to the cell surface.

Stieger, Bruno; Matter, Karl; Baur, Barbara; Bücher, K; Höchli, M.; Hauri, Hans Peter

doi:10.1083/jcb.106.6.1853

Cited by 59 publications

(66 citation statements)

References 40 publications

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“…In addition, a subapical cell compartment seems to function as a docking platform for vesicles containing functional proteins (98). Parental Caco-2 cells and clones have also been used to identify the mechanisms underlying the sorting and surface delivery of apical and basolateral proteins in human enterocytes (99)(100)(101)(102)(103)(104)(105)(106)(107) and to find out how functional intestinal proteins take their place in cell membrane domains, including brush border-associated functional proteins such as SI (82,84,86,90,94,100,106,(108)(109)(110)(111)(112)(113)(114), AP (110,111), lactasephlorizin hydrolase (108,115), maltase-glucoamylase (108), APN (90,108,111), DPP IV (82,90,100,108,(116)(117)(118)(119)(120)(121), angiotensin I-converting enzyme (108), ␣-glucosidase (122), p-aminobenzoic acid peptide hydrolase (108), SGLT1, GLUT1, GLUT2, GLUT3, and GLUT5 (123)(124)…”

Section: Differentiated Enterocyte-like Parental Caco-2 Cell Line Andmentioning

confidence: 99%

Pathogenesis of Human Enterovirulent Bacteria: Lessons from Cultured, Fully Differentiated Human Colon Cancer Cell Lines

Moal

Servin

2013

Microbiol Mol Biol Rev

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SUMMARY Hosts are protected from attack by potentially harmful enteric microorganisms, viruses, and parasites by the polarized fully differentiated epithelial cells that make up the epithelium, providing a physical and functional barrier. Enterovirulent bacteria interact with the epithelial polarized cells lining the intestinal barrier, and some invade the cells. A better understanding of the cross talk between enterovirulent bacteria and the polarized intestinal cells has resulted in the identification of essential enterovirulent bacterial structures and virulence gene products playing pivotal roles in pathogenesis. Cultured animal cell lines and cultured human nonintestinal, undifferentiated epithelial cells have been extensively used for understanding the mechanisms by which some human enterovirulent bacteria induce intestinal disorders. Human colon carcinoma cell lines which are able to express in culture the functional and structural characteristics of mature enterocytes and goblet cells have been established, mimicking structurally and functionally an intestinal epithelial barrier. Moreover, Caco-2-derived M-like cells have been established, mimicking the bacterial capture property of M cells of Peyer's patches. This review intends to analyze the cellular and molecular mechanisms of pathogenesis of human enterovirulent bacteria observed in infected cultured human colon carcinoma enterocyte-like HT-29 subpopulations, enterocyte-like Caco-2 and clone cells, the colonic T84 cell line, HT-29 mucus-secreting cell subpopulations, and Caco-2-derived M-like cells, including cell association, cell entry, intracellular lifestyle, structural lesions at the brush border, functional lesions in enterocytes and goblet cells, functional and structural lesions at the junctional domain, and host cellular defense responses.

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Section: Differentiated Enterocyte-like Parental Caco-2 Cell Line Andmentioning

confidence: 99%

Pathogenesis of Human Enterovirulent Bacteria: Lessons from Cultured, Fully Differentiated Human Colon Cancer Cell Lines

Moal

Servin

2013

Microbiol Mol Biol Rev

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“…Delivery of Ag 525 to the basolateral surface occurred 10-15 min after maturation in the Golgi (as assessed by endo H resistance) and was strictly polarized (50:1). The delivery of apical Ag (37), and similar halftimes of apical appearance have been reported for several hepatocyte and intestinal apical markers (11,39,40).…”

Section: Discussionmentioning

confidence: 92%

Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.

Bivic

Real

Rodríguez-Boulan

1989

Proc. Natl. Acad. Sci. U.S.A.

188

170

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We studied the surface delivery pathways followed by newly synthesized plasma membrane proteins in intestinal cells. To this end, we developed an assay and characterized an epithelial cell line (SK-CO-15) derived from human colon adenocarcinoma. Polarized confluent monolayers (2000 a cm2), grown on polycarbonate filter chambers, were pulsed with radioactive methionine/cysteine and, at different times of chase, the protein fraction reaching the apical or basolateral surface was recovered by domain-selective biotinylation, immunoprecipitation, and immobilized streptavidin precipitation. Both an apical and a basolateral marker were found to be delivered vectorially to the respective surface, with a sorting efficiency of 50:1 for the basolateral marker and 14:1 for the apical marker.The mechanisms by which polarized epithelial cells sort and address plasma membrane proteins to apical and basolateral domains are still poorly understood (1)(2)(3)(4)(5). Research in different model epithelial systems has delineated two possible biogenetic pathways. In the kidney cell line MDCK (Madin-Darby canine kidney), exogenous viral envelope proteins are segregated intracellularly in the trans Golgi network and vectorially delivered by transport vesicles to their respective plasma membrane domains (6-9). An endogenous MDCK marker, the basolateral Na+ ,K+-ATPase, is also vectorially delivered (10). On the other hand, metabolic labeling/cell fractionation experiments with rat hepatocytes (11) and native intestinal cells (12-14) have provided evidence for initial delivery of both apical and basolateral antigens to the basolateral domain, followed by sorting and relocation of the apical proteins. However, Danielsen and Cowell (15) reported the opposite conclusion (i.e., intracellular sorting) for apical intestinal aminopeptidase, in agreement with electron microscopic studies that failed to detect these enzymes on the basolateral surface (16-18). Given the uncertainties arising from the different methodologies and epithelial cell systems used, it is unclear to what extent intracellular sorting or sorting at the plasma membrane are responsible for the sorting ofapical and basolateral plasma membrane proteins in epithelial cells.Here we have used a different approach (metabolic labeling combined with domain-selective biotinylation and recovery) to study the delivery of endogenous apical and basolateral antigens in a new polarized human colon adenocarcinoma cell line. This procedure avoids subcellular fractionation procedures, which is difficult in cultured cells, and allows for simple and sensitive monitoring of the delivery of plasma membrane proteins from intracellular biogenetic pools to the respective plasma membrane domain.

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“…It is worth noting that the complex forms of dipeptidylpeptidase IV and sucrase-isomaltase were resistant to endoglycosidase H to the same extent as those of untreated control cells (data not shown). Moreover, forskolin treatment had no effect on the kinetics of appearance of dipeptidylpeptidase IV in the brush-border, as determined by pulse-chase experiments in conjunction with subcellular fractionation (Stieger et al, 1988; data not shown). Obviously, forskolin slightly modulates complex glycosylation in the Golgi apparatus by an unknown mechanism.…”

Section: Stimulation Of Lactase-phlorizin Hydrolase Biosynthesis By Fmentioning

confidence: 99%

“…Sucrase-isomaltase (Semenza, 1981(Semenza, , 1986Hauri, 1988) is also synthesized in Caco-2 cells as a 210-kDa high-mannose precursor form, undergoing complex glycosylation in the Golgi apparatus to a 217-kDa form, which is transported directly to the brush-border membrane (Hauri et al, 1985;Matter et al, 1990;Le Bivic et al, 1990). Unlike in the intestine under in vivo conditions (Hauri et al, 1979(Hauri et al, , 1980Sjostrom et al, 1980), sucrase-isomaltase is not cleaved into its sucrase and isomaltase subunits in Caco-2 cells due to the absence of pancreatic proteases in cell culture.…”

mentioning

confidence: 99%

Induction of lactase biosynthesis in the human intestinal epithelial cell line Caco‐2

Hauri

Sander

Naim

1994

European Journal of Biochemistry

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The human colonic adenocarcinoma cell line Caco-2 forms monolayers of differentiated enterocyte-like cells when cultured on permeable supports. After confluency, Caco-2 cells express a number of brush-border enzymes including lactase-phlorizin hydrolase, sucrase-isomaltase and dipeptidylpeptidase IV. We have studied, with particular emphasis on lactase-phlorizin hydrolase, the modulation of biosynthesis of these enzymes by stimulating second messenger systems. Forskolin induced lactase-phlorizin hydrolase synthesis approximately fourfold within 7 h, suppressed sucraseisomaltase synthesis, and had little effect on dipeptidylpeptidase IV. Dibutyryl-CAMP, 8-bromocAMP and vasoactive intestinal peptide also increased lactase-phlorizin hydrolase biosynthesis, indicating c-AMP dependent regulation. The induction of lactase-phlorizin hydrolase biosynthesis could be inhibited by actinomycin D and was preceded by a fourfold increase in lactase-phlorizin hydrolase mRNA levels, suggesting transcriptional control. Phorbol 12-myristate 13-acetate had an inhibitory effect on brush-border enzyme synthesis, in particular on sucrase-isomaltase, and blocked the forskolin-induced biosynthesis of lactase-phlorizin hydrolase. Lactase-phlorizin hydrolase synthesis was also inducible by hydrocortisone, but maximal induction required at least 3 days during which time sucrase-isomaltase synthesis diminished. The results indicate opposite regulation of lactase-phlorizin hydrolase and sucrase-isomaltase via cAMP and corticosteroids, and suggest that the Caco-2 cell line can serve as a model system to study aspects of the humoral regulation of human intestinal brush-border enzymes in cell culture.The human epithelial cell line Caco-2 has been established as a model system to study various aspects of intestinal epithelial cell biology including the synthesis of brushborder enzymes, protein trafficking, epithelial cell polarity (Zweibaum et al., 1991 ;Hauri and Matter, 1991 ;Louvard et al., 1992;Rodriguez-Boulan and Powell, 1992) and transepithelial transport (Thwaites et al., 1993). Although derived from a colonic adenocarcinoma, Caco-2 cells form monolayers of polarized cells on permeable supports and after confluency spontaneously express a number of features that are characteristic for differentiated small intestinal enterocytes rather than large intestinal colonocytes. As it has not been possible to establish non-transformed intestinal epithelial cell lines exhibiting a differentiated phenotype, Caco-2 remains the most promising cell line with which to study small intestinal functions in culture.Among other brush-border enzymes (Howell et al., 1992), Caco-2 cells express lactase-phlorizin hydrolase (LPH;Semenza, 1981Semenza, , 1986 Montgomery et al., 1991;Naim, 1993) which is synthesized as a 200-kDa high-man-

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Dissection of the asynchronous transport of intestinal microvillar hydrolases to the cell surface.

Cited by 59 publications

References 40 publications

Pathogenesis of Human Enterovirulent Bacteria: Lessons from Cultured, Fully Differentiated Human Colon Cancer Cell Lines

Pathogenesis of Human Enterovirulent Bacteria: Lessons from Cultured, Fully Differentiated Human Colon Cancer Cell Lines

Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.

Induction of lactase biosynthesis in the human intestinal epithelial cell line Caco‐2

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