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J. Biol. Chem., Vol. 281, Issue 27, 18269-18272, July 7, 2006
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Identification of the Maturation Factor for Dual Oxidase
EVOLUTION OF AN EUKARYOTIC OPERON EQUIVALENT*
1
¶
From the
Departments of Medicine, Pediatrics, and ¶Committee on Genetics, University of Chicago, Chicago, Illinois 60637
Received for publication, April 24, 2006
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ABSTRACT |
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 TOP ABSTRACT INTRODUCTIONEXPERIMENTAL PROCEDURESRESULTS AND DISCUSSIONREFERENCES |
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Dual oxidase 2 (DUOX2), an NADPH:O2 oxidoreductase flavoprotein, is a component of the thyroid H2O2 generator crucial for hormone synthesis at the apical membrane. Mutations in DUOX2 produce congenital hypothyroidism in humans. However, no functional DUOX-based NADPH oxidase has ever been reconstituted at the plasma membrane of transfected cells. It has been proposed that DUOX retention in the endoplasmatic reticulum (ER) of heterologous systems is due to the lack of an unidentified component required for functional maturation of the enzyme. By data mining of a massively parallel signature sequencing tissue expression data base, we identified an uncharacterized gene named DUOX maturation factor (DUOXA2) arranged head-to-head to and co-expressed with DUOX2. A paralog (DUOXA1) was similarly linked to DUOX1. The genomic rearrangement leading to linkage of ancient DUOX and DUOXA genes could be traced back before the divergence of echinoderms. We demonstrate that co-expression of DUOXA2, an ER-resident transmembrane protein, allows ER-to-Golgi transition, maturation, and translocation to the plasma membrane of functional DUOX2 in a heterologous system. The identification of DUOXA genes has important implications for studies of the molecular mechanisms controlling DUOX expression and the molecular genetics of congenital hypothyroidism.
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TOPABSTRACTINTRODUCTIONEXPERIMENTAL PROCEDURESRESULTS AND DISCUSSIONREFERENCES |
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Generation of H2O2 at the apical membrane of thyroid follicular cells is essential for iodination of thyroglobulin by thyroid peroxidase and constitutes the rate-limiting step of thyroid hormone synthesis. Dual oxidases (DUOX1 and DUOX2)2 appear to constitute the catalytic core of the H2O2 generator (1, 2). They are large homologs of the phagocyte gp91phox/Nox2 NADPH-dependent oxidase with an N-terminal extension comprising a peroxidase-like domain. Although the crucial role of DUOX2 in thyroid hormonogenesis has been substantiated by reports of severe congenital hypothyroidism in patients with biallelic nonsense mutations (3), the understanding of structure, function, and regulation of DUOX has remained limited. The major obstacle for molecular studies of DUOX is the lack of a suitable heterologous cell system for DUOX-based functional NADPH oxidase expression. Transfected cells completely retain DUOX in the endoplasmatic reticulum (ER) (4–8), suggesting that an unidentified component, essential for DUOX maturation, may be specifically expressed in tissues containing the functional enzyme.
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EXPERIMENTAL PROCEDURES |
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TOPABSTRACTINTRODUCTIONEXPERIMENTAL PROCEDURESRESULTS AND DISCUSSIONREFERENCES |
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Data Mining and Computational Analysis—Massively parallel signature sequencing (MPSS) data (9) were obtained from the NCBI Gene Expression Omnibus repository (www.ncbi.nlm.nih.gov/geo/; records GSE1747 and GPL1443). A thyroid specificity score, as defined by Jongeneel et al. (9), was calculated for signatures with frequency >100 tags per million (
30 mRNA copies/cell) in the thyroid/parathyroid library. Tags with scores >–1 were mapped to the human genome assembly using BLAST. DUOXA homologs were identified by tBLASTn searches against the NCBI nr data base and trace archive and BLAT queries (at genome.ucsc.edu/) against assembled whole genome sequences. Orthologs were operationally defined as reciprocal best BLAST hits. Gene structures were deduced by spliced alignment maintaining maximum homolog similarity of the open reading frames (ORFs) and consensus splice junctions. Cladograms were constructed from ClustalX alignments (BLOSUM weight matrix, excluding gaps) using the Jones, Taylor, and Thornton (JTT) substitution model in PHYML 2.4.4 (10). SignalP 3.0 (11) and Phobius (12) were used to analyze signal peptides, transmembrane helices, and topology. Northern Blot Analysis—A human multiple tissue Northern blot (Origene) was hybridized with DUOXA2 (125–470 of DQ489734) and DUOXA1 (1244–1623 of BC020841) probes.
Heterologous Expression of DUOX2 and DUOXA2 Constructs—cDNA was synthesized with Superscript reverse transcriptase (Invitrogen) by oligo(dT) priming of total RNA from a normal human thyroid gland. The DUOX2 and DUOXA2 ORFs were amplified using native Pfu polymerase (Stratagene) and cloned into pcDNA3.1 (Invitrogen). Epitope-tagged constructs and fusions with enhanced green fluorescent protein (EGFP) were prepared by replacement or splicing-by-overlap extension using specifically designed primers. All constructs were verified by sequencing. HeLa cells were cultured and transfected as described (13).
Confocal Laser Scanning Microscopy—Indirect immunofluorescence of permeabilized cells has been described previously (13). For surface staining, cells were incubated with rat anti-HA clone 3F10 and/or mouse anti-c-myc clone 9E10 (both from Roche Applied Science) at 1 µg/ml in Hank's buffered saline solution/10 mM Hepes, pH 7.4, 1% bovine serum albumin at 4 °C. Rabbit anti-calnexin was obtained from StressGen. Images were captured on a Nikon Eclipse E800 equipped with PCM2000.
Analysis of N-Glycosylation—Postnuclear supernatants (in 50 mM Tris/HCl, pH 8.0, 150 mM NaCl, and proteinase inhibitors) were adjusted to 0.5% SDS, 0.4 mM dithiotreitol and denatured, at room temperature, for 30 min. Samples were deglycosylated with N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) (both from New England Biolabs) according to manufacturer's recommendations, followed by SDS-PAGE under reducing conditions and Western blotting as described (13).
Measurement of H2O2 Generation—Release of H2O2 was determined by reaction with cell-impermeable 10-acetyl-3,7-dihydroxyphenoxazine (14) (Amplex Red reagent, Invitrogen) in the presence of excess peroxidase, producing fluorescent resorufin. Briefly, cell monolayers were incubated, with or without 10 µM diphenyleneiodonium (DPI), in Dulbecco's phosphate-buffered saline supplemented with 50 µM Amplex Red reagent and 0.1 unit/ml horseradish peroxidase for 1 h at 37°C. Relative fluorescence units (excitation/emission: 535/595) were corrected for Amplex Red oxidation in wells containing non-transfected cells and converted into H2O2 concentrations using a calibration curve. Renilla luciferase activity from co-transfected pRL-Tk plasmid (Promega) was used as internal control as described (13).
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RESULTS AND DISCUSSION |
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TOPABSTRACTINTRODUCTIONEXPERIMENTAL PROCEDURESRESULTS AND DISCUSSIONREFERENCES |
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Identification of Novel Genes in the DUOX1/DUOX2 Intergenic Region—We used MPSS data for 32 normal human tissues (9) to identify novel transcripts with predominant expression in thyroid gland. One of the extracted tags mapped to an uncharacterized locus (LOC405753) oriented head-to-head to DUOX2 in the
16-kbp DUOX1/DUOX2 intergenic region. For reasons outlined below, we called the corresponding gene DUOX maturation factor 2 (DUOXA2).3
Based on human-mouse homology (Riken clone 9030623N16Rik), and supported by contig assembly of expressed sequence tags (ESTs), it comprises six exons, confirmed by reverse transcription-PCR amplification from human thyroid tissue (GenBankTM accession number DQ489734). The putative transcription start site defined by clone DKFZp686C04213 maps to a GpC rich region (Fig. 1A). This site is 135 bp from the 5' terminus of a spliced DUOX2 EST (BI045475) on the opposite strand. A single polyadenylation signal (Fig. 1A) is supported by all mapped 3' ESTs. We confirmed a specific transcript of the expected size (1.3 kbp) by Northern blot analysis (Fig. 1B), which also validated the MPSS-based expression profiling: DUOXA2 mRNA was by far most abundant in thyroid, with lower levels in salivary glands reflecting the known expression profile of DUOX2 (1, 2, 15).
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The DUOXA2 ORF is initiated within a Kozak consensus (gcagcATGa) and spans all six exons. The encoded 320-amino acid protein was strongly predicted to comprise five membrane-integral regions, including a reverse signal anchor with external N terminus (type III) (Fig. 1C). The three NX(S/T) consensus sites for N-glycosylation are clustered within an extended external loop connecting the second and third transmembrane helices.
We identified a single DUOXA2 paralog in the human genome. We will refer to this locus, annotated as "homolog of Drosophila Numb-interacting protein," as DUOXA1. It is immediately adjacent, in tail-to-tail orientation to DUOXA2 and extends, via untranslated exons, into the DUOX1 promoter region. DUOXA1 mRNA was predominantly expressed in thyroid gland and, at lower level, in esophagus (Fig. 1B). Two transcripts of 2.9 and 3.5 kbp were detected, compatible with alternative splicing of 5'-untranslated exons and the use of alternative 3'-polyadenylation signals (data not shown). The DUOXA1 ORF was confirmed by sequencing from human thyroid cDNA (GenBankTM accession number DQ489735).
By spliced alignment, we deduced the gene structures of all DUOXA homologs in 10 other vertebrate whole genome assemblies. The splicing sites of all structures were conserved at the single codon level (exon alignment shown in Fig. 2S in the on-line supplement). Remarkably, the bidirectional DUOX/DUOXA arrangement was conserved throughout the vertebrate lineage (Fig. 1E, accession numbers of genomic contigs available in Table 4S in the on-line supplement). Teleosts have a single DUOX/DUOXA arrangement, which has undergone tandem duplication to an inverted repeat (DUOX2/DUOXA2/DUOXA1/DUOX1) before the amphibian divergence. Analyzing unassembled genomic contigs, we mapped the evolutionary event leading to the bidirectional association of DUOX and DUOXA before the divergence of echinoderms, since linkage of the loci was present in Strongylocentrotus purpuratus. Thus, conserved microsynteny in deuterostomes was a strong predictor for cooperation between DUOX and DUOXA.
The protostomes C. elegans and D. melanogaster lack a DUOXA homolog in the vicinity of their respective duox loci. They do, however, each harbor a single ancient DUOXA homolog. For instance, Drosophila moladietz (mol) encodes a 474-amino acid protein that exhibits 39% amino acid identity over 256 amino acids with human DUOXA1.
Functional Rescue of DUOX2 by DUOXA2—To test whether DUOXA2 can reconstitute DUOX2 activity in a heterologous system, we expressed either DUOX2, DUOXA2, or both in HeLa cells and measured H2O2 released into the culture medium. Transfection of either DUOX2 or DUOXA2 alone did not result in increased H2O2 generation compared with nontransfected cells, confirming previous results for DUOX2 (2, 4). Remarkably, co-transfection of DUOX2 with DUOXA2 rescued DUOX2 activity as indicated by the significant amounts of H2O2 released from the cells (Fig. 2A). The H2O2 release triggered by DUOX2/DUOXA2 co-transfection was completely blocked by the flavoprotein inhibitor DPI (Fig. 2A).
Co-expression of DUOXA2 Permits ER-Exit of DUOX2 and Plasma Membrane Targeting via the Secretory Pathway—Lack of DUOX2 activity in heterologous systems has been associated with absence of DUOX2 at the plasma membrane (4). To directly test whether reconstitution of active DUOX2 by DUOXA2 is indeed due to translocation of DUOX2 to the plasma membrane, we HA-tagged DUOX2 at its extracellular domain (HA-DUOX2; tag inserted between Asp27 and Ala28). Non-permeabilized cells showed strong anti-HA plasma membrane signals in cells co-transfected with HA-DUOX2 and DUOXA2 (Fig. 2B). Untransfected cells, or cells transfected with either DUOXA2 or HA-DUOX2 alone, were devoid of surface fluorescence (Fig. 2B and data not shown).
To determine whether DUOXA2-induced surface expression of DUOX2 involved ER-to-Golgi transition of DUOX2, we analyzed the maturation of DUOX2 N-glycan moieties using specific glycosidases. Whereas all N-glycans are cleavable by PNGase F, the ER-derived high-mannose type N-glycans become resistant to Endo H once they have been modified by Golgi-localized enzymes. HA-DUOX2 expressed in HeLa cells migrated as a single band on SDS-PAGE and was sensitive to full deglycosylation by Endo H, consistent with published data (4). In contrast, co-transfection with DUOXA2 resulted in the appearance of a second DUOX2 species with slightly decreased mobility and complete resistance to deglycosylation by Endo H (Fig. 2C). These findings resembled those previously obtained with endogenous DUOX2 protein (4, 5, 16), indicating that expression of DUOX2 in our reconstituted system involved normal maturation of DUOX2 within the secretory pathway.
Characterization of DUOXA2 as ER-resident Protein—DUOXA2 could be an integral part of a DUOX2 complex, endowing a holocomplex with the ability to exit the ER and reach the plasma membrane. We, therefore, determined whether myc-tagged DUOXA2 alone or in combination with DUOX2 would be detectable at the plasma membrane. Of several constructs tested, only DUOXA2 with N-(myc-DUOXA2) or C-terminal (DUOXA2-myc/His) attached myc tags were fully functional in rescuing DUOX2 activity as assessed by H2O2 generation and HA-DUOX2 plasma membrane targeting (data not shown). However, neither myc-DUOXA2 nor DUOXA2-myc/His was detectable at the plasma membrane (data not shown), although they had the expected size on Western blot analysis (Fig. 3A) and intracellularly co-localized with HA-DUOX2 (Fig. 3B).
To exclude that this was due to a discrepancy between the modeled and actual DUOXA2 membrane topology or due to masking of the N-terminal epitope tag, we fused EGFP/myc to the C terminus of DUOXA2 (an N-terminal fusion was not functional). As shown in Fig. 3C, DUOXA2-EGFP/myc did not co-localize with HA-DUOX2 at the plasma membrane, the latter delineated by anti-HA surface staining. The intracellular distribution of DUOXA2-EGFP/myc (and of DUOXA2-myc/His) showed a similar distribution pattern as the ER-marker calnexin (Fig. 3D).
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To further corroborate that DUOXA2 is indeed an ER-resident protein, we analyzed the maturation of DUOXA2-myc/His N-glycosylation in cells co-expressing HA-DUOX2. We found that the N-glycans of DUOXA2-myc/His were exclusively of the high-mannose type (Fig. 3E). In contrast, detection of HA-DUOX2 in the same samples demonstrated, again, that about half of DUOX2 protein had been subject to Golgi modification of its glycosylation (data not shown, compare Fig. 2C). Collectively, these results indicate that DUOXA2 is not an integral part of a DUOX2 enzyme complex at the plasma membrane but an ER-resident protein promoting ER exit and maturation of DUOX2. It should be noted that N-glycosylation of DUOXA2 supports our topology model (Fig. 1C), since the apparent molecular weight of the N-glycan moieties (
10 kDa) indicates N-glycosylation of all three consensus sites.
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An Evolutionary Perspective on the Physiological Role of DUOXA—The DUOX/DUOXA transcriptional unit is an excellent example of bidirectional transcription of tightly linked genes that are not structurally related but are involved in the same pathway, an arrangement considered equivalent to a prokaryotic operon (17). In contrast to the organization in deuterostomes, the two protostomal DUOXA-homologs analyzed were not genetically linked to duox; in fruit flies, there is also evidence for a distinct functional speciation. The Drosophila DUOXA-homolog (mol) encodes a plasma membrane protein (mol-PA) implicated in the polarized recruitment of a cytosolic signal mediator (Numb) to the plasma membrane (18). That mol-PA may not cooperate with Drosophila Duox is also suggested by the distinct phenotypes caused by deficiency of mol (memory loss; listed as CG4482 in Ref 19) or Drosophila duox (defective gut immunity) (20). Remarkably, this functional divergence relates to a distinct topology prediction of the region likely crucial for functional speciation of mol-PA and DUOXA. In DUOXA, the second and third transmembrane helices are connected by an extended luminal loop consistent with N-glycosylation of DUOXA2 (Figs. 1C and 3E). In mol-PA, the equivalent region harbors a binding motif crucial for recruitment of Numb (18), which, therefore, would have to be cytosolic. Analysis of the residue-wise posterior probabilities for a given state (inside/outside/transmembrane) in the 1-best Phobius topology models indeed supports such a scenario of distinct membrane topologies: the profile of transmembrane probabilities in mol-PA, but not vertebrate DUOXA, indicates the potential for two additional membrane-spanning helices (Fig. 1D), which would flank the Numb-binding motif resulting in its cytosolic exposure.
If, as we propose, DUOX expression in Drosophila does not require mol, what could have been the advantage of a DUOX/DUOXA system that caused its maintenance in deuterostomes over more than 500 million years (21) of divergent evolution? For unexplained reasons, in sea urchin eggs (22) and likewise in follicular thyroid cells (4), the bulk of DUOX protein is not detected at the cell surface but in intracellular compartments, which could provide a stimulus-recruitable pool. Thus, from an evolutionary perspective, the emergence of DUOXA may have provided an additional level of DUOX regulation, specifically, the control of DUOX translocation to the plasma membrane. The ability to reconstitute active DUOX enzyme will provide the tool to investigate the molecular mechanisms underlying DUOX expression in diverse model systems.
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FOOTNOTES |
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* This work was supported in part by National Institutes of Health Grants DK15070 and DK20595. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1S–3S and Table 4S.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) DQ489734 and DQ489735.
1 To whom correspondence should be addressed: Dept. of Medicine, The University of Chicago, 5841 S. Maryland Ave., MC3090, Chicago, IL 60637. Tel.: 773-702-9273; Fax: 773-702-6940; E-mail: hgrasber{at}uchicago.edu.
2 The abbreviations used are: DUOX, dual (domain) oxidase; BLAST, basic local alignment search tool; DPI, diphenyleneiodonium; EGFP, enhanced green fluorescent protein; ER, endoplasmatic reticulum; EST, expressed sequence tag; HA, hemagglutinin; MPSS, massively parallel signature sequencing; ORF, open reading frame; PNGase F, N-glycosidase F; Endo H, endoglycosidase H; contig, group of overlapping clones.
3 The gene name and symbol have been approved by the HUGO Gene Nomenclature Committee.
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REFERENCES |
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TOPABSTRACTINTRODUCTIONEXPERIMENTAL PROCEDURESRESULTS AND DISCUSSIONREFERENCES |
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- Dupuy, C., Ohayon, R., Valent, A., Noel-Hudson, M. S., Deme, D., and Virion, A. (1999) J. Biol. Chem. 274, 37265–37269
[Abstract/Free Full Text] - De Deken, X., Wang, D., Many, M. C., Costagliola, S., Libert, F., Vassart, G., Dumont, J. E., and Miot, F. (2000) J. Biol. Chem. 275, 23227–23233
[Abstract/Free Full Text] - Moreno, J. C., Bikker, H., Kempers, M. J., van Trotsenburg, A. S., Baas, F., de Vijlder, J. J., Vulsma, T., and Ris-Stalpers, C. (2002) N. Engl. J. Med. 347, 95–102
[Abstract/Free Full Text] - De Deken, X., Wang, D., Dumont, J. E., and Miot, F. (2002) Exp. Cell Res. 273, 187–196[CrossRef][Medline] [Order article via Infotrieve]
- Morand, S., Chaaraoui, M., Kaniewski, J., Deme, D., Ohayon, R., Noel-Hudson, M. S., Virion, A., and Dupuy, C. (2003) Endocrinology 144, 1241–1248
[Abstract/Free Full Text] - Morand, S., Agnandji, D., Noel-Hudson, M. S., Nicolas, V., Buisson, S., Macon-Lemaitre, L., Gnidehou, S., Kaniewski, J., Ohayon, R., Virion, A., and Dupuy, C. (2004) J. Biol. Chem. 279, 30244–30251
[Abstract/Free Full Text] - Wang, D., De Deken, X., Milenkovic, M., Song, Y., Pirson, I., Dumont, J. E., and Miot, F. (2005) J. Biol. Chem. 280, 3096–3103
[Abstract/Free Full Text] - Ameziane-El-Hassani, R., Morand, S., Boucher, J. L., Frapart, Y. M., Apostolou, D., Agnandji, D., Gnidehou, S., Ohayon, R., Noel-Hudson, M. S., Francon, J., Lalaoui, K., Virion, A., and Dupuy, C. (2005) J. Biol. Chem. 280, 30046–30054
[Abstract/Free Full Text] - Jongeneel, C. V., Delorenzi, M., Iseli, C., Zhou, D., Haudenschild, C. D., Khrebtukova, I., Kuznetsov, D., Stevenson, B. J., Strausberg, R. L., Simpson, A. J., and Vasicek, T. J. (2005) Genome Res 15, 1007–1014
[Abstract/Free Full Text] - Guindon, S., and Gascuel, O. (2003) Syst. Biol. 52, 696–704[CrossRef][Medline] [Order article via Infotrieve]
- Bendtsen, J. D., Nielsen, H., von Heijne, G., and Brunak, S. (2004) J. Mol. Biol. 340, 783–795[CrossRef][Medline] [Order article via Infotrieve]
- Kall, L., Krogh, A., and Sonnhammer, E. L. (2004) J. Mol. Biol. 338, 1027–1036[CrossRef][Medline] [Order article via Infotrieve]
- Grasberger, H., Ringkananont, U., Lefrancois, P., Abramowicz, M., Vassart, G., and Refetoff, S. (2005) Mol. Endocrinol. 19, 1779–1791
[Abstract/Free Full Text] - Zhou, M., Diwu, Z., Panchuk-Voloshina, N., and Haugland, R. P. (1997) Anal. Biochem. 253, 162–168[CrossRef][Medline] [Order article via Infotrieve]
- Geiszt, M., Witta, J., Baffi, J., Lekstrom, K., and Leto, T. L. (2003) FASEB J. 17, 1502–1504
[Abstract/Free Full Text] - El Hassani, R. A., Benfares, N., Caillou, B., Talbot, M., Sabourin, J. C., Belotte, V., Morand, S., Gnidehou, S., Agnandji, D., Ohayon, R., Kaniewski, J., Noel-Hudson, M. S., Bidart, J. M., Schlumberger, M., Virion, A., and Dupuy, C. (2005) Am. J. Physiol. 288, G933–G942
- Gavalas, A., Dixon, J. E., Brayton, K. A., and Zalkin, H. (1993) Mol. Cell. Biol. 13, 4784–4792
[Abstract/Free Full Text] - Qin, H., Percival-Smith, A., Li, C., Jia, C. Y., Gloor, G., and Li, S. S. (2004) J. Biol. Chem. 279, 11304–11312
[Abstract/Free Full Text] - Dubnau, J., Chiang, A. S., Grady, L., Barditch, J., Gossweiler, S., McNeil, J., Smith, P., Buldoc, F., Scott, R., Certa, U., Broger, C., and Tully, T. (2003) Curr. Biol. 13, 286–296[CrossRef][Medline] [Order article via Infotrieve]
- Ha, E. M., Oh, C. T., Bae, Y. S., and Lee, W. J. (2005) Science 310, 847–850
[Abstract/Free Full Text] - Douzery, E. J., Snell, E. A., Bapteste, E., Delsuc, F., and Philippe, H. (2004) Proc. Natl. Acad. Sci. U. S. A. 101, 15386–15391
[Abstract/Free Full Text] - Wong, J. L., Creton, R., and Wessel, G. M. (2004) Dev. Cell 7, 801–814[CrossRef][Medline] [Order article via Infotrieve]
- Pachucki, J., Wang, D., Christophe, D., and Miot, F. (2004) Mol. Cell. Endocrinol. 214, 53–62[CrossRef][Medline] [Order article via Infotrieve]
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