The Receptor Tyrosine Kinase Ror2 Associates with the Melanoma-associated Antigen (MAGE) Family Protein Dlxin-1 and Regulates Its Intracellular Distribution

The mammalian Ror family receptor tyrosine kinases, Ror1 and Ror2, play crucial roles in developmental morphogenesis. Although the functions of Ror1 and Ror2 are redundant, Ror2 exhibits more specific functions during development. We show that when expressed in mammalian cells, Ror2, but not Ror1, associates with the melanoma-associated antigen (MAGE) family protein, Dlxin-1, which is known to bind to the homeodomain proteins Msx2 and Dlx5 and regulate their transcriptional functions. This association requires the cytoplasmic C-terminal region of Ror2, containing proline-rich and serine/threonine-rich domains, and the C-terminal necdin homology domain of Dlxin-1. Interestingly, the cytoplasmic C-terminal region of Ror2 is missing in patients with brachydactyly type B. Interestingly, transient expression and immunohistochemical analyses reveal that both Dlxin-1 and Msx2 are co-localized in the nuclei in the absence of Ror2. In the presence of Ror2, Dlxin-1 is co-localized with Ror2 at the membranous compartments and Msx2 is retained in the nuclei. It was also found that the majority of cellular Dlxin-1 is retained in the membrane fractions of wild-type but not Ror2 –/– mouse embryonic fibroblasts. Furthermore, we show that transcriptional activity of Msx2, irrespective of Ror2 kinase activity, is regulated by ectopic expression of Ror2 using a reporter plasmid containing the WIP element. Thus, Ror2 sequesters Dlxin-1 in membranous compartments, thereby affecting the transcriptional function of Msx2. The mammalian Ror family receptor tyrosine kinases, Ror1 and Ror2, play crucial roles in developmental morphogenesis. Although the functions of Ror1 and Ror2 are redundant, Ror2 exhibits more specific functions during development. We show that when expressed in mammalian cells, Ror2, but not Ror1, associates with the melanoma-associated antigen (MAGE) family protein, Dlxin-1, which is known to bind to the homeodomain proteins Msx2 and Dlx5 and regulate their transcriptional functions. This association requires the cytoplasmic C-terminal region of Ror2, containing proline-rich and serine/threonine-rich domains, and the C-terminal necdin homology domain of Dlxin-1. Interestingly, the cytoplasmic C-terminal region of Ror2 is missing in patients with brachydactyly type B. Interestingly, transient expression and immunohistochemical analyses reveal that both Dlxin-1 and Msx2 are co-localized in the nuclei in the absence of Ror2. In the presence of Ror2, Dlxin-1 is co-localized with Ror2 at the membranous compartments and Msx2 is retained in the nuclei. It was also found that the majority of cellular Dlxin-1 is retained in the membrane fractions of wild-type but not Ror2 –/– mouse embryonic fibroblasts. Furthermore, we show that transcriptional activity of Msx2, irrespective of Ror2 kinase activity, is regulated by ectopic expression of Ror2 using a reporter plasmid containing the WIP element. Thus, Ror2 sequesters Dlxin-1 in membranous compartments, thereby affecting the transcriptional function of Msx2. Receptor tyrosine kinases (RTKs) 1The abbreviations used are: RTK, receptor tyrosine kinase; BDB, brachydactyly type B; MAGE, melanoma-associated antigen; NRAGE, neurotrophin receptor-interacting MAGE homologue; NTR, neurotrophin receptor; NHD, necdin homology domain; HA, hemagglutinin; WT, wild type; MEF, mouse embryonic fibroblast; E, embryonic day (e.g. E12.5); HEK, human embryonic kidney; FCS, fetal calf serum; PBS, phosphate-buffered saline; DK, kinase dead.1The abbreviations used are: RTK, receptor tyrosine kinase; BDB, brachydactyly type B; MAGE, melanoma-associated antigen; NRAGE, neurotrophin receptor-interacting MAGE homologue; NTR, neurotrophin receptor; NHD, necdin homology domain; HA, hemagglutinin; WT, wild type; MEF, mouse embryonic fibroblast; E, embryonic day (e.g. E12.5); HEK, human embryonic kidney; FCS, fetal calf serum; PBS, phosphate-buffered saline; DK, kinase dead. play crucial roles in developmental morphogenesis by regulating cellular proliferation, differentiation, migration, and death (1Schlessinger J. Cell. 2000; 103: 211-225Abstract Full Text Full Text PDF PubMed Scopus (3456) Google Scholar). The Ror family of RTKs are orphan RTKs, characterized by the presence of extracellular Frizzled-like cysteine-rich domains, membrane-proximal Kringle domains, and intracellular distal proline-rich domains that are assumed to mediate protein-protein interactions (2Forrester W.C. Cell. Mol. Life Sci. 2002; 59: 83-96Crossref PubMed Scopus (90) Google Scholar, 3Masiakowski P. Carroll R.D. J. Biol. Chem. 1992; 267: 26181-26190Abstract Full Text PDF PubMed Google Scholar, 4Masiakowski P. Yancopoulos G.D. Curr. Biol. 1998; 8: R407Abstract Full Text Full Text PDF PubMed Google Scholar, 5McKay S.E. Hislop J. Scott D. Bulloch A.G.M. Kaczmarek L.K. Carew T.J. Sossin W.S. Mol. Cell. Neurosci. 2001; 17: 821-841Crossref PubMed Scopus (26) Google Scholar, 6Oishi I. Sugiyama S. Liu Z.-J. Yamamura H. Nishida Y. Minami Y. J. Biol. 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Terashima T. Takada S. Yamamura H. Akira S. Minami Y. Genes Cells. 2000; 5: 71-78Crossref PubMed Scopus (190) Google Scholar). Mice lacking Ror2 exhibit dwarfism, short limbs (with mesomelic dysplasia) and tails, facial abnormalities, ventricular septal defects, and respiratory dysfunction resulting in neonatal lethality (12DeChiara T.M. Kimble R.B. Poueymirou W.T. Rojas J. Masiakowski P. Valenzuela D.M. Yancopoulos G.D. Nat. Genet. 2000; 24: 271-274Crossref PubMed Scopus (196) Google Scholar, 14Takeuchi S. Takeda K. Oishi I. Nomi M. Ikeya M. Itoh K. Tamura S. Ueda T. Hatta T. Otani H. Terashima T. Takada S. Yamamura H. Akira S. Minami Y. Genes Cells. 2000; 5: 71-78Crossref PubMed Scopus (190) Google Scholar). Ror1-deficient mice also die soon after birth due to respiratory dysfunction, yet they do not show any apparent skeletal or cardiac phenotypes (13Nomi M. Oishi I. Kani S. Suzuki H. Matsuda T. Yoda A. Kitamura M. Itoh K. Takeuchi S. Takeda K. Akira S. Ikeya M. Takada S. Minami Y. Mol. Cell. Biol. 2001; 21: 8329-8335Crossref PubMed Scopus (108) Google Scholar). Furthermore, Ror1/Ror2 double mutant mice exhibit markedly enhanced skeletal and cardiac abnormalities compared with Ror2 mutant mice, indicating that Ror1 and Ror2 interact genetically and functionally during the development of these organs (13Nomi M. Oishi I. Kani S. Suzuki H. Matsuda T. Yoda A. Kitamura M. Itoh K. Takeuchi S. Takeda K. Akira S. Ikeya M. Takada S. Minami Y. Mol. Cell. Biol. 2001; 21: 8329-8335Crossref PubMed Scopus (108) Google Scholar). These findings demonstrate the pleiotropic and specific, yet partially redundant, functions of Ror2 in mouse development. Interestingly, it has recently been reported that mutations within Ror2 are responsible for brachydactyly type B (BDB), a dominant skeletal disorder characterized by hypoplasia/aplasia of distal phalanges (15Oldridge M. Fortuna A.M. Maringa M. Propping P. Mansour S. Pollitt C. DeChiara T.M. Kimble R.B. Valenzuela D.M. 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Genet. 2000; 25: PubMed Scopus Google in This the crucial function of Ror2 in developmental morphogenesis. is known the by Ror1 and Ror2. was to proteins that interact with Ror1 Ror2, and a Dlxin-1, a of the melanoma-associated antigen (MAGE) family Y. S. A. T. H. Itoh K. PubMed Scopus Google Scholar, K. M. T. K. Biochem. PubMed Scopus Google Scholar, P. C. P. C. E. van B. A. T. PubMed Scopus Google Scholar), was Dlxin-1 has been to with the homeodomain proteins Msx2 and Dlx5 and to regulate the transcriptional function of Dlx5 Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). The of Dlxin-1, or has also been characterized B. E. A. M. M. P. J. G. 2000; PubMed Scopus Google Scholar, C. A. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). has been to bind to the neurotrophin receptor and to C. A. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). In it has been reported that with of and of the D. J. R. M. S. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). Furthermore, it has recently been that and a a and S. J. Biol. Chem. 2002; Full Text Full Text PDF PubMed Scopus Google Scholar). we show that Ror2, but not Ror1, associates with Dlxin-1 when expressed in mammalian The association of Ror2 with Dlxin-1 requires the cytoplasmic C-terminal region of Ror2, which containing proline-rich and serine/threonine-rich This region is in Ror2 a of Dlxin-1 in the of Interestingly, Dlxin-1 and Msx2 are co-localized in the nuclei in the absence of Ror2, the of Dlxin-1, but not Msx2, is by the presence of Ror2. These that Dlxin-1 is important of Ror2 function and that is regulated in by Ror2. Ror1 and Ror2 I. Takeuchi S. Hashimoto R. Nagabukuro A. Ueda T. Liu Z.-J. Hatta T. Akira S. Matsuda Y. Yamamura H. Otani H. Minami Y. Genes Cells. 1999; 4: 41-56Crossref PubMed Scopus (110) Google Scholar). the of the the cytoplasmic domains of Ror1 and Ror2 the or Ror proteins The to Ror1 and Ror2 by using a of specific that for Ror1 and for Ror2, at the of these and the C-terminal region of Ror2, containing the proline-rich and serine/threonine-rich domains, containing the and serine/threonine-rich domains and the of the kinase The expression a mutant of Ror2, was by crucial for with In the expression Ror2 or Ror2 and compared with by expression not The expression and Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). The reporter plasmid was H. G. H. P. N. C. Mol. Cell. Biol. 1997; 17: PubMed Scopus Google Scholar). and the and the the human was used to mouse Dlxin-1 A. Y. K. K. K. J. Biol. Chem. 2002; Full Text Full Text PDF PubMed Scopus Google Scholar). and and and embryonic using a H. J. Biol. 17: PubMed Scopus Google Scholar). and in with fetal calf was using the Y. Y. A. T. K. Yamamura H. T. Full Text PDF PubMed Scopus Google Scholar), or to the was the the and by the the cytoplasmic of of mouse Ror1 and Ror2, at the of was with the or with the mouse in the plasmid for their to lacking The for activity by and and when and and found to the containing the that Dlxin-1 protein, by with Life by with the plasmid by a of and with and and by at for of fractions type and Ror2 was Y. J. R.D. J. Biol. Chem. Full Text PDF PubMed Google Scholar). and for at with The with to for at The with of the and with The or by and to membrane The with the and with to or using Life I. Sugiyama S. Liu Z.-J. Yamamura H. Nishida Y. Minami Y. J. Biol. Chem. 1997; 272: 11916-11923Abstract Full Text Full Text PDF PubMed Scopus (83) Google Scholar, I. Sugiyama S. Otani H. Yamamura H. Nishida Y. Minami Y. Mech. Dev. 1998; PubMed Scopus Google Scholar). by in for at and with containing for at in with for with in PBS, for at with and with in PBS, at for in PBS, the with and with with the reporter plasmid H. G. H. P. N. C. Mol. Cell. Biol. 1997; 17: PubMed Scopus Google with the expression by using The of was with of of expression plasmid was in and to using the The in In analyses Y. T. Takada S. Dev. Biol. 1997; PubMed Scopus Google Scholar). for Ror1, Ror2, and T. Nomi M. Ikeya M. Kani S. Oishi I. Terashima T. Takada S. Minami Y. Mech. Dev. 2001; 105: 153-156Crossref PubMed Scopus (115) Google Scholar, Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). of Dlxin-1 a the roles of Ror1 and Ror2 during we a to cytoplasmic that with the cytoplasmic of Ror1 and Ror2. mouse was using the mouse cytoplasmic of Ror1 and Ror2 We Dlxin-1, a of the MAGE a Ror1 and Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google not Dlxin-1 associates with Ror1 Ror2 in mammalian cells, Dlxin-1 and Ror1 or Ror2 in in Dlxin-1 with Ror2, but not with Ror1, indicating that Ror2, but not Ror1, associates with Dlxin-1 in We or not association of Ror2 with Dlxin-1 requires Ror2 kinase Dlxin-1 with wild-type or a Ror2 mutant Ror2 expressed in cells, and their association was by by in Ror2 WT, but not Ror2 DK, was tyrosine when in by of It was found that both Ror2 and Ror2 with Dlxin-1 to a indicating that association of Ror2 kinase It has been reported that a of the Ror family RTKs, important roles in regulating cellular migration, of and of during development. It has also been reported that the tyrosine kinase activity of is for of these functions but not for in cellular (2Forrester W.C. Cell. Mol. Life Sci. 2002; 59: 83-96Crossref PubMed Scopus (90) Google Scholar, W.C. M. E. G. 1999; PubMed Scopus (134) Google Scholar). Thus, it is that Dlxin-1 may mediate the tyrosine of Ror2. of Ror2 and Dlxin-1 during the of the association in cells, we compared the embryonic expression patterns of Ror1, Ror2, and Dlxin-1 by in analyses mouse at and not Ror2 and Dlxin-1 expression in the and limb Ror1 transcripts a of Ror2 and Dlxin-1 expressed the limbs, Ror1 expression was more to the of the limb the expression patterns of these transcripts in the a Dlxin-1, Ror2, was expressed in the of the and the of the expression was at 11Matsuda T. Nomi M. Ikeya M. Kani S. Oishi I. Terashima T. Takada S. Minami Y. Mech. Dev. 2001; 105: 153-156Crossref PubMed Scopus (115) Google not the Ror1 transcripts detected in the and of the limbs 11Matsuda T. Nomi M. Ikeya M. Kani S. Oishi I. Terashima T. Takada S. Minami Y. Mech. Dev. 2001; 105: 153-156Crossref PubMed Scopus (115) Google not The reveal that the expression patterns of Ror2 and Dlxin-1 overlap in the and This that Ror2 with Dlxin-1 during mouse The C-terminal of Ror2 and the C-terminal of Dlxin-1 for the of Ror2 with a within Ror2 that is for association with Dlxin-1, we of Ror2 and Ror2 the C-terminal containing the and domains, containing the and domains and of the kinase We the of these to with Dlxin-1 in in Ror2 with Dlxin-1, indicating that the C-terminal region of Ror2, containing the and domains, is for Although the cytoplasmic tyrosine kinase domains of Ror1 and Ror2 exhibit of their C-terminal exhibit a of of It also be that Ror2 patients exhibits and mutations that a of Ror2 lacking the C-terminal proline-rich and serine/threonine-rich domains or the cytoplasmic region (15Oldridge M. Fortuna A.M. Maringa M. Propping P. Mansour S. Pollitt C. DeChiara T.M. Kimble R.B. Valenzuela D.M. Yancopoulos G.D. Wilkie A.O.M. Nat. Genet. 2000; 24: 275-278Crossref PubMed Scopus (177) Google Scholar, 16Schwabe G.C. Tinschert S. Buschow C. Meinecke P. Wolff G. Gillessen-Kaesbach G. Oldridge M. Wilkie A.O.M. Kömec R. Mundlos S. Am. J. Hum. Genet. 2000; 67: 822-831Abstract Full Text Full Text PDF PubMed Scopus (134) Google Scholar). It has been reported that associates with the of Dlxin-1 the cytoplasmic domain of C. A. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). apparent was found the cytoplasmic C-terminal region of Ror2 and the cytoplasmic domain of It be of to Ror2 and for association with Dlxin-1 in Dlxin-1 to the MAGE family of characterized by the presence of in their C-terminal Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google and C. A. 2000; Full Text Full Text PDF PubMed Scopus Google is of the characterized proteins in and has been to with and and the function of during H. N. M. K. J. Biol. Chem. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar, H. K. K. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar). It that the is in protein-protein In the region of Dlxin-1 protein, is a of the which is to of with Dlx5 Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, The domain of Dlxin-1 not show homology to any in the We of Dlxin-1 for their association with Ror2, in We found that Dlxin-1 but not Dlxin-1 or Dlxin-1 with Ror2. This that the of Dlxin-1 is responsible for association with Ror2. the and the of Dlxin-1 for with Dlxin-1 was a protein, it was that Dlxin-1 associates with and Msx2 Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). Dlxin-1 associates with Msx2 more with Dlx5 or family Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, not We which of Dlxin-1 is responsible for with Msx2. Msx2 and Dlxin-1 Dlxin-1 or Dlxin-1 expressed in cells, and their association was by by in Dlxin-1 and Dlxin-1 but not Dlxin-1 with Msx2, indicating that both the domain and the of Dlxin-1 are for association with Msx2. the that the of Dlxin-1 is for association with both Ror2 and Msx2, it was assumed that the association of Ror2 and Msx2 with Dlxin-1 is we found that Msx2 was not with Ror2 in the presence of Dlxin-1 the the association Ror2 and Dlxin-1 was by the absence or presence of Msx2 This is with Msx2 and Ror2 for Dlxin-1, that the Ror2 and Dlxin-1 to the that it any Msx2 and Dlxin-1. of Dlxin-1 by Msx2 and the of the of Ror2 and Msx2 with Dlxin-1, we the of Ror2, Dlxin-1, and of Msx2 proteins in by Ror2, Dlxin-1, and Msx2 expressed in cells, Ror2 was found to be in the membranous compartments the membrane and Dlxin-1 and Msx2 in the and We found that Dlxin-1 was co-localized with Ror2 or Msx2 when expressed in B and This is with analyses and Dlxin-1 is co-localized with Msx2 in the nuclei Dlxin-1 is co-localized with Ror2 at the membrane and the when Ror2 and Msx2 in cells, Ror2 and Msx2 proteins detected at the membranous compartments and in the and their intracellular not overlap at when expressed in cells, was detected at the membrane and detected in the membranous compartments and in the nuclei These that Dlxin-1 is co-localized with Ror2, but not with Msx2 in the membranous compartments, Msx2 is retained in the nuclei. This is with analyses that Ror2 and Msx2 with Dlxin-1, with Ror2 of Dlxin-1 with Ror2, but not with Msx2, was by a using Dlxin-1, Ror2, and Msx2 expressed in not The Ror2 Ror2 to Dlxin-1 in Ror2 mutant Ror2 to with Dlxin-1 in a We Ror2 is co-localized with Dlxin-1. was expressed by in cells, it at the membrane and to wild-type Ror2 Ror2 and Dlxin-1 Ror2 was detected in the membranous compartments, Dlxin-1 the when Ror2 and Msx2 in cells, both proteins detected in the membranous compartments and in the and their intracellular overlap at Dlxin-1, Ror2 and Msx2 in cells, was detected in the nuclei Ror2 Msx2 and This that Dlxin-1 with Msx2 in the Ror2 in the membranous of Dlxin-1 with Msx2, but not Ror2 was also by a using Dlxin-1, Ror2 and Msx2 in not these that the Ror2 Ror2 to Dlxin-1 in the membranous Ror2 mutant more cytoplasmic Ror2 also to Dlxin-1 in the membranous compartments not of Dlxin-1 by that Ror2 sequesters Dlxin-1 in the membranous compartments, we of the wild-type and Ror2 –/– mice Because mouse Ror2 are the presence or absence of Ror2 expression in the wild-type and Ror2 –/– was by and analyses not of Dlxin-1 not the wild-type and Ror2 –/– not that expression of Dlxin-1 is by Ror2. in the majority of cellular Dlxin-1 was in the membrane fractions in wild-type with in the the in Ror2 –/– a of cellular Dlxin-1 was detected in the membrane the of Dlxin-1 was Ror2 –/– and wild-type The of cellular Dlxin-1 was also Ror2 –/– and wild-type The that a of cellular Dlxin-1 in Ror2 –/– may be in the This that Ror2 is responsible for the of Dlxin-1 and that it sequesters Dlxin-1 in the membrane of Msx2 by of the of of Dlxin-1 by Ror2 in the membranous compartments, we expression of Ror1 WT, Ror2 WT, or Ror2 the transcriptional activity of Msx2. with with the expression with a reporter plasmid containing a WIP of the and a reporter H. G. H. P. N. C. Mol. Cell. Biol. 1997; 17: PubMed Scopus Google Scholar). ectopic of Dlxin-1, it was found that Dlxin-1 is expressed in at a not with a H. G. H. P. N. C. Mol. Cell. Biol. 1997; 17: PubMed Scopus Google Scholar), of the Msx2 expression in transcriptional of the WIP reporter plasmid in transcriptional by Msx2 was at least partially by ectopic expression of Ror2 but not Ror1 Interestingly, it was also at least partially by ectopic expression of Ror2 that Ror2, irrespective of kinase activity, the transcriptional activity of Msx2 in the nuclei by Dlxin-1, a transcriptional for Msx2, in the membranous Dlxin-1 a of Ror2 the Ror family RTKs of structurally related Ror1 and Ror2 P. Carroll R.D. J. Biol. Chem. 1992; 267: 26181-26190Abstract Full Text PDF PubMed Google Scholar, 7Oishi I. Takeuchi S. Hashimoto R. Nagabukuro A. Ueda T. Liu Z.-J. Hatta T. Akira S. Matsuda Y. Yamamura H. Otani H. Minami Y. Genes Cells. 1999; 4: 41-56Crossref PubMed Scopus (110) Google Scholar). Although both Ror1 and Ror2 play crucial roles during Ror2 exhibits more specific functions compared with Ror1 (13Nomi M. Oishi I. Kani S. Suzuki H. Matsuda T. Yoda A. Kitamura M. Itoh K. Takeuchi S. Takeda K. Akira S. Ikeya M. Takada S. Minami Y. Mol. Cell. Biol. 2001; 21: 8329-8335Crossref PubMed Scopus (108) Google Scholar). We have that Dlxin-1, a of the MAGE family of associates with Ror2, but not with Ror1 This that Dlxin-1 may be in not of the developmental functions of Ror2. Interestingly, Ror2 and Dlxin-1 exhibit expression patterns in the and limbs, in to Ror1, which a This that the association Ror2 and Dlxin-1 may be during development. Furthermore, it has been that the cytoplasmic C-terminal region of Ror2, containing proline-rich and serine/threonine-rich domains, and the C-terminal necdin homology domain of Dlxin-1 are for the association Ror2 and Dlxin-1 and It also be that the cytoplasmic C-terminal region of Ror2 is missing in a of Dlxin-1 in the of Dlxin-1 is co-localized with the wild-type Ror2 in the membranous compartments the of by Ror2 and the mutant Ror2 to Dlxin-1 in the membranous compartments the functions of the MAGE family of proteins for the was to mediate of by with and K. M. T. K. Biochem. PubMed Scopus Google Scholar, H. N. M. K. J. Biol. Chem. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar, H. K. K. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar). Interestingly, it has recently been that Dlxin-1 is a Dlxin-1 associates with the homeodomain proteins Msx2 and Dlx5 and the transcriptional function of the Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google associates with and is for of C. A. 2000; Full Text Full Text PDF PubMed Scopus Google and associates with the and and the of D. J. R. M. S. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). to that Ror2 and Msx2 bind to region within Dlxin-1 and it has been reported that and association with are also C. A. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). findings the roles of Ror2 during development. Dlxin-1 has been to with and regulate the transcriptional activity of Dlx5 Y. A. H. N. K. K. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar), a of the family of homeodomain proteins in skeletal development. it is that the transcriptional activity of Msx2, homeodomain in skeletal development I. H. M. I. K. T. Y. M. S. H. R. R. Nat. Genet. 2000; 24: PubMed Scopus Google Scholar, A.O.M. Elanko N. S. Nat. Genet. 2000; 24: PubMed Scopus Google Scholar), be by with Dlxin-1. In we show that transcriptional of the WIP reporter plasmid by Msx2 was partially by ectopic of Ror2 and Ror2 DK, The the that of Dlxin-1 by Ror2 at the membranous compartments, irrespective of kinase activity, the transcriptional function of Msx2 in the nuclei. Furthermore, it is that of Ror2 by may the of Dlxin-1. the expression of Ror2, Dlxin-1, and of Msx2 in a particular may be the that regulate the and function of the or Although the of Ror2 is may the regulating the and function of Dlxin-1 Msx2. We M. for a of the We also K. for