Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
J Physiol
2019 Jan 01;5972:521-542. doi: 10.1113/JP276714.
Show Gene links
Show Anatomy links
Anticipation of food intake induces phosphorylation switch to regulate basolateral amino acid transporter LAT4 (SLC43A2) function.
Oparija L
,
Rajendran A
,
Poncet N
,
Verrey F
.
???displayArticle.abstract???
KEY POINTS: Amino acid absorption requires luminal uptake into and subsequent basolateral efflux out of epithelial cells, with the latter step being critical to regulate the intracellular concentration of the amino acids. The basolateral essential neutral amino acid uniporter LAT4 (SLC43A2) has been suggested to drive the net efflux of non-essential and cationic amino acids via parallel amino acid antiporters by recycling some of their substrates; its deletion has been shown to cause defective postnatal growth and death in mice. Here we test the regulatory function of LAT4 phosphorylation sites by mimicking their phosphorylated and dephosphorylated states in Xenopus laevis oocytes and show that dephosphorylation of S274 and phosphorylation of S297 increase LAT4 membrane localization and function. Using new phosphorylation site-specific antibodies, we observe changes in LAT4 phosphorylation in mouse small intestine that correspond to its upregulation at the expected feeding time. These results strongly suggest that LAT4 phosphorylation participates in the regulation of transepithelial amino acid absorption.
ABSTRACT: The essential amino acid uniporters LAT4 and TAT1 are located at the basolateral side of intestinal and kidney epithelial cells and their transport function has been suggested to control the transepithelial (re)absorption of neutral and possibly also cationic amino acids. Uniporter LAT4 selectively transports the branched chain amino acids leucine, isoleucine and valine, and additionally methionine and phenylalanine. Its deletion leads to a postnatal growth failure and early death in mice. Since LAT4 has been reported to be phosphorylated in vivo, we hypothesized that phosphorylation regulates its function. Using Xenopus laevis oocytes, we tested the impact of LAT4 phosphorylation at Ser274 and Ser297 by expressing mutant constructs mimicking phosphorylated and dephosphorylated states. We then investigated the in vivo regulation of LAT4 in mouse small intestine using new phosphorylation site-specific antibodies and a time-restricted diet. In Xenopus oocytes, mimicking non-phosphorylation of Ser274 led to an increase in affinity and apparent surface membrane localization of LAT4, stimulating its transport activity, while the same mutation of Ser297 decreased LAT4's apparent surface expression and transport rate. In wild-type mice, LAT4 phosphorylation on Ser274 was uniform at the beginning of the inactive phase (ZT0). In contrast, at the beginning of the active phase (ZT12), corresponding to the anticipated feeding time, Ser274 phosphorylation was decreased and restricted to relatively large patches of cells, while Ser297 phosphorylation was increased. We conclude that phosphorylation of small intestinal LAT4 is under food-entrained circadian control, leading presumably to an upregulation of LAT4 function at the anticipated feeding time.
Albrecht,
Timing to perfection: the biology of central and peripheral circadian clocks.
2012, Pubmed
Albrecht,
Timing to perfection: the biology of central and peripheral circadian clocks.
2012,
Pubmed
Amanchy,
A curated compendium of phosphorylation motifs.
2007,
Pubmed
Antalis,
Mechanisms of disease: protease functions in intestinal mucosal pathobiology.
2007,
Pubmed
Balakrishnan,
Circadian clock genes and implications for intestinal nutrient uptake.
2012,
Pubmed
Bedet,
Constitutive phosphorylation of the vesicular inhibitory amino acid transporter in rat central nervous system.
2000,
Pubmed
Bodoy,
Identification of LAT4, a novel amino acid transporter with system L activity.
2005,
Pubmed
,
Xenbase
Bröer,
Amino acid homeostasis and signalling in mammalian cells and organisms.
2017,
Pubmed
Bröer,
Amino acid transport across mammalian intestinal and renal epithelia.
2008,
Pubmed
Böhmer,
Characterization of mouse amino acid transporter B0AT1 (slc6a19).
2005,
Pubmed
,
Xenbase
Chen,
Cyclic GMP kinase II (cGKII) inhibits NHE3 by altering its trafficking and phosphorylating NHE3 at three required sites: identification of a multifunctional phosphorylation site.
2015,
Pubmed
Christensen,
Quantitative phosphoproteomics dissection of seven-transmembrane receptor signaling using full and biased agonists.
2010,
Pubmed
Damiola,
Restricted feeding uncouples circadian oscillators in peripheral tissues from the central pacemaker in the suprachiasmatic nucleus.
2000,
Pubmed
Dave,
Expression of heteromeric amino acid transporters along the murine intestine.
2004,
Pubmed
Davies,
Mutations of the BRAF gene in human cancer.
2002,
Pubmed
Deribe,
Post-translational modifications in signal integration.
2010,
Pubmed
Gazzola,
The adaptive regulation of amino acid transport system A is associated to changes in ATA2 expression.
2001,
Pubmed
Gilbert,
Board-invited review: Peptide absorption and utilization: Implications for animal nutrition and health.
2008,
Pubmed
González,
Regulation of the neuronal glutamate transporter excitatory amino acid carrier-1 (EAAC1) by different protein kinase C subtypes.
2002,
Pubmed
Guetg,
Essential amino acid transporter Lat4 (Slc43a2) is required for mouse development.
2015,
Pubmed
,
Xenbase
Havel,
Peripheral signals conveying metabolic information to the brain: short-term and long-term regulation of food intake and energy homeostasis.
2001,
Pubmed
Hediger,
The ABCs of solute carriers: physiological, pathological and therapeutic implications of human membrane transport proteinsIntroduction.
2004,
Pubmed
Hornbeck,
PhosphoSitePlus, 2014: mutations, PTMs and recalibrations.
2015,
Pubmed
Horák,
The role of ubiquitin in down-regulation and intracellular sorting of membrane proteins: insights from yeast.
2003,
Pubmed
Hundal,
Amino acid transceptors: gate keepers of nutrient exchange and regulators of nutrient signaling.
2009,
Pubmed
Hussain,
Circadian Regulation of Macronutrient Absorption.
2015,
Pubmed
Jando,
Expression and regulation of the neutral amino acid transporter B0AT1 in rat small intestine.
2017,
Pubmed
Karasov,
Adaptive regulation of sugar and amino acid transport by vertebrate intestine.
1983,
Pubmed
Karve,
Small changes huge impact: the role of protein posttranslational modifications in cellular homeostasis and disease.
2011,
Pubmed
Kim,
Systematic and quantitative assessment of the ubiquitin-modified proteome.
2011,
Pubmed
Konturek,
Gut clock: implication of circadian rhythms in the gastrointestinal tract.
2011,
Pubmed
Liu,
Gastrin decreases Na+,K+-ATPase activity via a PI 3-kinase- and PKC-dependent pathway in human renal proximal tubule cells.
2016,
Pubmed
Maciejewski,
Mutation of serine 90 to glutamic acid mimics phosphorylation of bovine prolactin.
1995,
Pubmed
Makrides,
Transport of amino acids in the kidney.
2014,
Pubmed
Mariotta,
T-type amino acid transporter TAT1 (Slc16a10) is essential for extracellular aromatic amino acid homeostasis control.
2012,
Pubmed
McGivan,
Regulatory and molecular aspects of mammalian amino acid transport.
1994,
Pubmed
Meier,
Activation of system L heterodimeric amino acid exchangers by intracellular substrates.
2002,
Pubmed
,
Xenbase
Mertins,
Integrated proteomic analysis of post-translational modifications by serial enrichment.
2013,
Pubmed
Mizoguchi,
Human cystinuria-related transporter: localization and functional characterization.
2001,
Pubmed
,
Xenbase
Mohawk,
Central and peripheral circadian clocks in mammals.
2012,
Pubmed
Moolenbeek,
The "Swiss roll": a simple technique for histological studies of the rodent intestine.
1981,
Pubmed
Nguyen,
When ubiquitination meets phosphorylation: a systems biology perspective of EGFR/MAPK signalling.
2013,
Pubmed
Nässl,
Amino acid absorption and homeostasis in mice lacking the intestinal peptide transporter PEPT1.
2011,
Pubmed
Palacín,
Molecular biology of mammalian plasma membrane amino acid transporters.
1998,
Pubmed
Pan,
Diurnal rhythm of H+-peptide cotransporter in rat small intestine.
2002,
Pubmed
Pan,
The diurnal rhythm of the intestinal transporters SGLT1 and PEPT1 is regulated by the feeding conditions in rats.
2004,
Pubmed
Pendergast,
Robust food anticipatory activity in BMAL1-deficient mice.
2009,
Pubmed
Poncet,
The role of amino acid transporters in nutrition.
2013,
Pubmed
Pácha,
Circadian regulation of epithelial functions in the intestine.
2013,
Pubmed
Ramadan,
Recycling of aromatic amino acids via TAT1 allows efflux of neutral amino acids via LAT2-4F2hc exchanger.
2007,
Pubmed
,
Xenbase
Ramadan,
Basolateral aromatic amino acid transporter TAT1 (Slc16a10) functions as an efflux pathway.
2006,
Pubmed
,
Xenbase
Reischl,
Kinases and phosphatases in the mammalian circadian clock.
2011,
Pubmed
Robles,
Phosphorylation Is a Central Mechanism for Circadian Control of Metabolism and Physiology.
2017,
Pubmed
Rossier,
LAT2, a new basolateral 4F2hc/CD98-associated amino acid transporter of kidney and intestine.
1999,
Pubmed
,
Xenbase
Rouillard,
The harmonizome: a collection of processed datasets gathered to serve and mine knowledge about genes and proteins.
2016,
Pubmed
Saito,
Regulatory mechanism governing the diurnal rhythm of intestinal H+/peptide cotransporter 1 (PEPT1).
2008,
Pubmed
Samluk,
Protein kinase C regulates amino acid transporter ATB(0,+).
2012,
Pubmed
Schibler,
Clock-Talk: Interactions between Central and Peripheral Circadian Oscillators in Mammals.
2015,
Pubmed
Schneider,
NIH Image to ImageJ: 25 years of image analysis.
2012,
Pubmed
Shacter,
Energy consumption in a cyclic phosphorylation/dephosphorylation cascade.
1984,
Pubmed
Silk,
Protein digestion and amino acid and peptide absorption.
1985,
Pubmed
Stephan,
Entrainment of circadian rhythms by feeding schedules in rats with suprachiasmatic lesions.
1979,
Pubmed
Szabó,
Phosphorylation site mutations in the human multidrug transporter modulate its drug-stimulated ATPase activity.
1997,
Pubmed
Tanaka,
Functional expression and adaptive regulation of Na+ -dependent neutral amino acid transporter SNAT2/ATA2 in normal human astrocytes under amino acid starved condition.
2005,
Pubmed
Taslimifar,
Quantifying the relative contributions of different solute carriers to aggregate substrate transport.
2017,
Pubmed
,
Xenbase
Venne,
The next level of complexity: crosstalk of posttranslational modifications.
2014,
Pubmed
Verrey,
System L: heteromeric exchangers of large, neutral amino acids involved in directional transport.
2003,
Pubmed
Vilches,
Cooperation of Antiporter LAT2/CD98hc with Uniporter TAT1 for Renal Reabsorption of Neutral Amino Acids.
2018,
Pubmed
Wagner,
Proteomic analyses reveal divergent ubiquitylation site patterns in murine tissues.
2012,
Pubmed
Welsh,
Suprachiasmatic nucleus: cell autonomy and network properties.
2010,
Pubmed
Wu,
Dietary protein intake and human health.
2016,
Pubmed