Encoding Structural of Epidermal Cornification and S100 Calcium-Binding Proteins Form a Complex ("Epidermal Differentiation Complex") on lq21

Dietmar Mischke,* Bernhard P. Korge,t Ingo Marenholz,* Armin Volz,* and Andreas Ziegler*

*Insti tut flir Experimente llc Onkologie und Transplantationsmedizin, Virchow-Kliniku111 der Hutllbold t-Univc rsitiit ZlI Berl in. Berlin; and t Klinik lind Poliklinik fUr Demlatologic und Venerologie, Universitii t Z lI Knln. Knln. Gemlan),

Chromosome 1 reveals in region lq21 a most remark­ chohyalin, profilaggrin, , SPRR3, SPRRIB, able density of genes that fulfill important functions SPRR2A, , S100A9, StOOA8, StOOA6. Co­ in terminal differentiation of the human . localization of genes expressed late during matura­ These genes encode the cornified envelope precur­ tion of epidermal cells together with genes encodin g sors loricrin, involucrin, and small proline-rich pro­ calcium-binding proteins is particularly intriguing teins (SPRRt, SPRR2, and SPRR3), the intermediate since calciulll levels tightly control the differe ntia ­ filament-associated proteins profilaggrin and tri­ tion of epithelial cells and the expression of genes chohyalin, and several S100A calciUln-binding pro­ encoding epidermal structural proteins. Accounting teins. Extending and refining our previous physical for the close functional cooperation alllong the se tnap of lq21 we have now mapped two additional structurally and evolutionary related genes, we con ­ S100A genes as well as the three SPRR subfamilies clude that these loci constitute a gene cOlllplex, for and resolved the arrangement of involucrin, SPRRs, which we propose the name epidermal differentiation and loricrin. All genes are linked within 1.9 Mbp of complex. ] llll'cst D CI'lrrato/ 106:989-992, 1996 human genomic DNA in the order: S100AI0, tri-

hromosome 1, which is the largest human chromo­ proteins contain 1:\>,1 0 calcium-binding dom ains with an EF-hand some and harbors about 8% of the entire human m oti f and share both a high degree of homology at the genomic information, reveals in region 1q21 a most level and a strikingly similar gene o rganizatio n. Since these genes remarkable clustering of gen es that fulfill important carried a pletho ra o f different names, a new logical no m en clature functions in epidermal differentiation. As judged (Sl OOA 1-S1 OOA I O) has recently been suggested and approved by frCom and gene structural information as w ell as evolution­ the Genome D ata Base [5]. T he third gene f.'l mily located in 1q21 ary relationships, these genes belong to three f.'lmilies and en code includes the multifuncti onal pro fil aggrin and tricho hyalin gen es. p roteins involved in the fonnation of the cornified envelope T hese are apparently " fu sed" genes containing two calcium­ (CE) of the human epidermis, S100A calcium-binding proteins, and binding m otifs like those in the S100A f.'l l1lily fo Howed by tandem intermediate filament-associated proteins [Ref. 1 and references peptide repeats that are characteristi c of the fa mily of CE genes th e rein]. [6,7,8]. M all Y of the 1q21 genes are also potential can didate gen es T he family of CE precursor genes includes loricrin, involucrin, for inherited disorders o f keratinization , such as tile large and and the small proline-rich proteins (SPRRs). The latter have been h eterogen eous gro ups of ichthyoti c disorders and psoriasis (9) . further subdivided into three classes consisting of two SPRR1 In this paper w e present an ex tended and refin ed hig h-resolution gen es, eight SPRR2 genes, and one SPRR3 gene [2]. Apart from a physical m ap of the 1 q21 region resolving the m ap positio n s of 11 very similar gene structure in terms of exon-intron distribution, the loci within 1.9 Mbp of geno mic D N A, including the fo u r newly CE precursors contain a central region of short tandem pepti de m apped genes S100A8, S100A9, SPRR2A. and SPR R3. Likewise, rep eats. T h e number of repeats vari es from 39 fo r involucrin, 20 fo r the positions of involucrin and SPR.Rl B relative to each other and loricrin, co 3-14 repeats for the different SPRRs [3]. T he second to the additio nal SP iU~s has been unambiguously detenn ined. group of genes residing in lq21 encompasses calcium-binding proteins of the S1 00A family [1 ,4,5, and this report]. All S1 00A MATERIALS AND METHODS DNA Probes The 3'-noncoding 17R-bp clone pL30.8 - 2 [1 01 and the Manuscrip t received N ovember 27 , 199 5; accepted for publica tion 6-kbp genomic clone p,\1-3 H6B PI] we re uscd to probe fnr 101;crin and January 2, 1996. involucrin. respecti vely. as previousl), described [1]. For SPRRIB. SP I ~R2A. R eprint rcqucsts to: Dr. Dietmar Mischke, institut flir Experimentelle and SPRR3 thc inserts of clone pJ C-I SB (470 bp) 112], p930 -1 (680 bp) Onkologic und Transplantationsmedizill, Virchow-Klinikul11 , Spandauer [1 2], and pSPR3 (320 bp) [2]. respcctively. were ava il abl e. Probes specific Damm 130, 0 -14050 Berlin, German),. for thc Sl OOA8 and S100A9 gcncs [1 3] were generated b)' pol),merase chain Abbreviations: CE, comifted cell cnvelope; EDC, epidermal differentia­ reaction ampliftcation using prim crs in the 3' -noncoding rcgion to ),ield a tion complex; SPRR, small proline rich protein. 60 1-bp S100A8 probe (upper prim cr: S'-TCTTATGCTTTTGTGGAAT_

002 2-202X/96/S1 0.50 • Copyright © 1996 by The Society for In vestigative Dermatology. Inc.

989 990 MISC HKE E1' AI- TH E JOUrtNAL OF I NVESTIGATI VE D ErtMATOLOGY

GAGGTT-3'; lower prim er: 5'-TGAGTTGGGAGTTTGGGAGTGGA-3') and a 402-bp S I OOA9 probe (upper primer: 5'-GCCACCCTGCCTCTAC­ CCA-3 ' ; lower primer: 5'-TGCCTTTTTGCTTTCTACAGTG-3'). Poly­ Illerase chain rea ction was performed using standard reaction conditions for 32 cycles (95°C, 60 s; 60°C, 60 s; 72°C. 120 s) . The initial denaturation was 5 min at 95 °C. and the fina l extension was 7 min at 72°C. Southern Blotting Agarosc blocks were prepared from the Epstein-Barr virus-tran sfo rm ed normal B-ccll lin e H2LCL. Aliquots co ntaining 5 X 10 5 -740kb cell s were digested overlli ght with 30 ullits of various rare cutting restriction -270kb -670kb enzymes as described previously [1 ). Fragments were separated by rotating -210kb fie ld gel electroph oresis [14 ] usin g the fo llowing pa rameters on the n..otaphor; duration. 36 h; interval (pulse time), '10-130 s lincar; angle. 125-1000 lin ear; field strength. 5.7-6.7 V/ cm linear; temperature. 13°C; agarose. 0.9'Yo, Scakem Generic Technology Grade (FMC Bioproducts. R.ockland. ME). After 10 min of HCI depurination. DNA was alkaline 90kb transferred to a nylon membrane (GeneScreen Plus; Dupont/ New England Nuclear, Boston, MA) and UV linked. Probes were labeled with J2p using the Megaprime kit (Amcrsham Buchler, Braunschweig. Germany) . South­ ern hybridization was done as described previously [I]. and bound probe was detected by autorad iography using Kodak XAR. fi lm .

RESULTS AND DISCUSS[O N La rge restri ction fra gm ents of human genomic DNA (of the Epstein-Barr virus-transformed normal B-cell line H2LCL) o b­ tained by sing le or double digestion with rare cutting enzym es w ere separated by rotating field gel electrophoresis, bl otted, and hybrid­ - 70kb ized sequen tia ll y w ith gene-specific probes. R epresentative results from these experiments are shown in Fig 1. T hei r interpretation takes advantage of the two chrom osom al 1 q21 haplotypes of H2LCL that differ from each other, probably because of genetic polymorphism and/or differential methylati on in the respective restriction sites. For example, probes for loricrin, S100A8 (formerl y calg ran ulin A (MRP8), CAGA), and S100A9 (formerl y calgranulin B (MRP14), CAGB) hybridized to the sam e 270- kbp Sjil fragment of one hapl otype, whereas the other revealed co-locali zation of on.ly the two SlOOA genes on a 210-kbp fragment (Fig ia). T he .S100A genes were resolved by SIft digestion , yielding a common 740-kbp fragm ent in o ne haplotype but two separate fragments of 670 and 70 kbp for S100A9 and S100A8, respectively, in the other -230kb (Fig lb). Similarl y, the order ofinvolucrin and th e three subfamjlies of SPRR genes could be determined (Fig 1c), thus resolving the ambiguity of o ur previo us map [1]. C learl y, the involucrin probe -180kb hybridized to a different Sail fragment (230 kbp) than any of three SPlU~ probes, allowing its unequivocal positioning on the restri c­ ,... ·tiII ;·,·1 - 140kb tion map. While SPRR3 and SPR.R1 B shared a 140-kbp Sail fragment, their order was definitely determined through the Salll - 110kb Sjil double d igest. Si nce the SPRR2A probe hybridized to yet another Sail fi'agment of 170 kbp, this gene could also be locali zed, thereby confirming previous m apping data for th e SPRJl... genes [2]. Finall y, the orientation of the SPRR llluitigene family within the 1q21 cluster co uld be settled unambig uo usly by li nking it physically to its neighbors, involucri n :lI1d loricrin. T he excended and improved physical m ap of the lq21 region is presented in Fig 2. Compared with o ur previous physical m ap [1] we have now m apped a total of 11 loci w ithin 1.9 Mbp and retined the extensions and distances of the correspon din g genes. At large, the members of the three gene fa milies seem to be grouped togeth er like the intermediate filament-associated genes trieh ohya­ Figure 1. Hybridization of gene-specific DNA probes to whole lin and pro fil aggrin or the CE precursOl' genes involucrin, 5Plill3, genome restriction fragments separated by rotating field gel elec­ SPR.RIB, SPR. .R 2A, and loricrin. T his ho lds true also for the SlOOA trophoresis. Autoradiob'Tams of individual lancs arc shown containing genes locH ted at o ne end of the map, except for only on e gene genomic DNA di gested with the indicated restriction cnzymes 'JIld hybrid­ (S 1 OOA 10; formerl y calpactin l li gh t ch ail1 , CALI L) that is posi­ izcd with gene-specifi c DNA probcs as indicated. Fil ters were re-used after tioned 1,500 kbp away, at the opposite border of tbe region. As stripping with 0. 1 X salin e sodiulll citrate and I % sodium dodecyl sul fate at the orientation of Sl OOA8 and 5i00A9 w ith respect to each other 99 °C for 10 min . All post-hybridization was hings were can;cd out under condi tions of high stringcncy (O.2 X salinc sodium citrate, 65 °C) . Sizes were and to 5100A6 (formerly calcydin, CACY) is in complete agree­ calculated fro III a co-electrophoresed size marker. IVL, involucrin; LOR., ment w ith data reported for a yeast artifi ci:d chrom osome clone loricJi Il. containing nine S1 OOA genes within approximately 350 kbp [5], we incorporated this information in to Fig 2 to link six additional Co-localization of genes withi.n the appears to be S100A genes to o ur m ap. A total of 25 genes have now been rather fi·cquent. Yet, two differcnt situati ons may be distinguished. phys icall y locali zed to this region of 1 q21, considering the multi­ O ne is the case of gene cl usters, i.e., the locali zation of genes within plicity of som e SPRR genes [2]. a close distance that share a high degree of homology and there- VOL. 106, NO. 5 MA Y 1996 EPID ERMAL DIFFER£NTIATION COMPLEX IN Iq21 991

I I Olm 500kb 1000kb 2000kb

~ q,.o ~' o~ ,p 1\ 10..0 . ~ ~ !\~10.. . ~' 10..~' '1\ ~ .~' ~, ~"'~'l>0~,~!i.~ ~, ~'O"" 0" ~' «f'" ~I '2:i<0 ~~'" 0 !i.~ ~v ~~v '

Srfl II, I II ~I I, II ~I, .1 I BssHIi II I, I II II ~I' I .1 I Sfil II I' I 1·1 1'1 II' ... 1.1' I -I ., I ~I' I -1-. Sail I II I II I' ' I' 'i' I -I I I II II I Srfl II, I II I, II III II -II I BssHIi II I, I II I II , I, I -I~ II II II _, I Sfil II I I, I 1, 1 I, .1, I _I, I 4 I I ~I_I II --' Sail II I II II 14 _II ,.. I I _I I III II II I I I- ~ 5\'\ .~' . ~',p \ ~v 'O~'s- \ ~ <0"" <0"''<0'''''' \ \

\ \ --- q, \ v-'O ;. \ oV- 00 ov-~v-%vr:,;> o-?Y oV-" \ ,,0 00 ,,0 "," 0~~~~a~"o "'.... '" = DO~D 0 '"

Figure 2. Rotatil1g field gel electrophoresis restric tion map of the epidermal differel1tiatiol1 complex il1 lq21. The linc" ,. arrangement of the mapped genes is shown with respect to the two hapl otypes present in the 1-1 2LCL cell line as establi shed w ith the rare cutting restriction enzymcs SrJT , Bssi-lIl . sfir, and Snll. T hi s map is supported by restriction analys is wich further enz ymes (Bsi\Xl I. NO lI. M llIl, N,."I. and Snell ) as reported previo usly IlJ . Considering tbe two S p~n..'I, eight SPIU:l..2 . and onc SPIU:l..3 genes [2) as well as the other Sl OOA genes id entified on a yeast artificial clone IS), the EDC harbors at least 25 genes. T l-lI-I, : FLG. profilaggrin; IVL. involu cl-il1 ; LOR. loricrin .

fore belong to the sa me gene famil y. T he other con cept regards process of epidermal differentiation and tightly control the ;11 1/ ;/('0 co-locali zation as a m eans through w hich gene complexes will be expression of at Least the profil aggrin and lo ri crin genes [1 8,19]. created that unite functionall y interacting genes belonging to Accordingly. all of these genes arc potential candidate genes tor different gene famili es. Examples for gene clusters are the type I and inherited disorders of keratinization and many of them have already type II keratin genes on chromosom es 17q and 12q, respectively been implicated with such diseases. For example, highly aberrant [15]. or the immunoglobuli n heavy- chain cluster on chro m osom e expression of profilaggrin and abnormal CEs have been descl; bed 14q [16]. The most prominent example for a gene complex is th e in ichthyotic disorders [20,21]. Also. SlOOA7, S100A8, and S100A9 major hi stocompatibility (HLA) complex on chro mosome 6p [1 7J (formerly psoriasill (PSO.R 1). calgranuli n A (MRP8), and calgranu­ harboring a large number o f functional genes, of which many arc lin B (MRP 14), respectively) h ave been shown to be highly crucial for the normal function of the immune system . Som e upregul ated in psoriatic epidermis and primary human keratino­ e ncode the highly polymorphic HLA class I and class IT m o lecules cytes undergoing abno rmal differentiation [22,23J. Furthermore, that are involved in antigen presentation, w hil e other, unrelated, m ost recently, the autosom al recessive disorder ofpycnodysostosis, gen es encode either proteins responsible fo r the in tracellular pro­ which is characterized by sclerosing skeletal dysplasia and aero­ cessing of antigens or the transport of pep tides to be presented by osteolysis of distal phalanges, has been m apped to lq21, and the HLA class 1 molecules. S 1 OOA genes were considered pl ausible candidate genes [24,25]. Accordingly, the close physica l linkage .in lq2 'J of genes belong­ H ence, informatio n co nceming the chrom osom al organization of ing to different gene fam ilies together with their functional in ter­ EDC genes in 1 q21 will be of great va lue for gen etic Jink~ ge dep endence during cornification of the human epidem1is suggests analyses in a fFected families and for unraveling the underlYll1 g to u s that these genes also constitute a gene complex. Because this defects. complex is centrall y involved in the process of terminal epidermal T he functio nal similarities between the S100A proteins and the differentiation, it merits design ation as an epidermal differentiation calcium-binding in termediate fi lament-associated proteins as well complex (EDC). as m any other gene- and protein-structural ti es among all three Notably, in the fmal steps of kerati nocyte differentiation that gen e f.'lmi li es within the EDC m ay be more than coincidental. T his provide the human skin w ith a protective barrier against the raises questions that have likewise been addressed iJ1 the context of environment, several EDC-encoded gene products, such as lori­ the HLA complex, namely, the evolutionary history and biological erin, involucrin, SPRRs, and possibly other, not yet identified role o f the EDC (e.g., are the genes kept together simply by precursors, are cross-Linked by to form the highly accident or are there fu nctional constraints in keeping genes in close insoluble CEo At the sam e time, profilaggrin is processed to phys ical distance over millions ofyeaJ'S and across species barriers?). filaggrin, w hich leads to aggregation of keratin fi laments inside In this regard, analysis o f th e a -globin and the HLA class II loci has cornified cells. The co-localization of epidermal structural genes shown that co-locali zation of genes can signi fy the existen ce of a together with calcium-binding genes is even m ore intriguing locus control region, whose dominan t presence is necessary for be cause calcium levels are cri tically involved in coordinating the optimal and perhaps is al so of regulatory and 992 MISCH KE ET AL THE J OURNAL OF INVESTIGATIVE DERMATOLOGY transcriptional advantage to the linked genes [26,27]. In analogy, 9. R oop 0: Defects in the harrier. Srit·lIft· 267:474-475. 1995 som e genes of the EDC m ay also share superior locus control 10. Hohl D. Mehrel T. Li chti U. Turner ML. R oop OR, Swin er< PM: C haracter­ ization of lluman ioricrill . j Hi'" C hclII 266:6626-6636. \ 99 '\ regulatory elements coordinating their tissue- and differentiation­ t 'l . Eckert R. Green H : Structure and evolu tion of lhe humall illvolucrin gene. Cell specifi c transcription. Furthermore, analysis of the EDC throughout 46:583- 589. 1986 vertebrate evolution should al so shed Li ght on the functional 12. K:trtasova T. van del" Puttc P: Isolation, charactcriznliol1 ;md UV-srimularcd importance of linbge within the epidermal differentiation com­ express ion of two f.1 l11iH cs of genes encoding po lypeptides of related structure plex. [n this respect it is notew orthy that the mouse loricrin and in hum;ll1 epidcrl11al kcratilloc)' tcs. l\tIol Cdl Bioi 8:2 195-2203. 1988 13. Lagasse E. C lerc RG: C loning; and expression of twO human genes encoding pro fil aggrin genes are also linked [28] and that the pericentromeri c calcium-binding proteins th.lt arc rcgul nred during TIl )'cioid diUcrenriatio ll . J\1ol region of both arms of human has been found to be Cell BioI 8:2402- 24 10. 1988 syntenic with a continuous ' segm ent of mouse chromosome 3 14. Ziegler A. Volz A: Rotating field gel electrophoresis (ROFE). IH clJ/ilds J\'IlI i Bioi defining a putative homologous linkage group [29]. We have 12:63-72. 1992 therefore initiated a yeast artificial chromosome clone-based clon­ 15. Yoon 5J . Lel3lanc-StraceskiJ. Ward D. Kr;.uter K. Kucherl apari R: O rgani zation of the human keratin type II J;e ll c dustcr.ll 12'1 13. Gc,wlI/;o· 19:502- S0S. 1 99~ in g strategy for in depth analysis of the entire human EDC and to I G. Cook GP. Tomlinson 1M , Walter G. R..i ethmnn 1-1. Carter NP. Buluwela L. address som e of the questions raised above. Winter G . R..abbitts TI-I : A map of the human illlIllulloglobulin VH locus complcted by analysis of lhc telolllcric region of d m) l1l osoJl1 e 14'1. Nm Cellci 7: 162-168. 1994 17. M.i lncr C M . C'llnphcll R.D: Gelles, gcncs and m ore genes in the human maj or ~VC Ihllll k Drs. Ko zo )' 011 ('(10 olldJoseph emwll jor Ih" lorierill mill illl,,,llIedll DNA histocompatibility complex. Bim'ss(1)'s 14:565-57 1. 1992 c/olles, respec/i l,e/y, 01111 Drs. TOllio Km10sollo olld Clollde Bockelld"'Ijor specific 18. I-Iohl D, Lichti U, Breitkreutz D. Steinert PM. R oop DR: T ranscription of 'he prol)cs 10 ih e 111I"l"I' c/osses oj SPRR gCll es. This ",ork ",os slIpporl cd by 0 .grn llt j,..,,11 hum;m lo ri crin genc i" 1/ ;1 1'11 is induccd by calcium nnd cell density alld Ihe DClltsehe Forsc/lIIlIgsgcllleillschoji 10 Bel"llI/O,..1 P. Korge olld Diellllor Mischke. suppressed by retinoic Olein . ) 1"llcSI Dl'nllt1lo/ 96:4 14 -418. 1991 19. Yllspa SI-I, Kilkcnny AE. Steinert PM. Roop DR: Ex pression of murine epidermal differentiation 1ll00rkel's is ti ghliy regulated by restricted cxtracellular REFER E NCES ca lci um COll cclltrntions i" " jll'O .) Cdl Billl l 09: 1207-12 17. 19M9 20. Resin g KA, Dale BA: Proteins of keratohyalin. In: Goldsmith LA (cd.). 1. Volz A. Korge BIl. Compton JG. Ziegler A. Steinert PM, Mischke 0: Phys ical IJ/I),sio/flS}'. IJiodu.'lIIiJ'r}" (/ml AI/lllc{I//t,,. Biology (If ,I'I' Skill . Oxford Universiry mapping of a function:!l cluster of epidermal differentiation genes 0 11 chromo­ Press. N ew York, 1991. PI' 148-1 (,7 some Iq2 1. Cwo",;cs 18:92-99. 1993 21. Michels S. Juhlin L: Corniticd envelopcs in congcniwi disorders ofkcratini zadon. 2. Gihbs S. Fijllcl1l anll R. W icgant J, vn n Kess el AG. van de PUti C P, 1l;l ckclldorfC: Br) Omllato/ 122:IS- 2 1. 19YO Molectllar characterization and evolution of the ~PRR. f.1 lllil y of kcratinocytc 22. Hoffmann HJ . O lscn E, Etzcrodt M . Mndscn P. Thogerscn H e . Kruse T. Celis ditTerentiation lI1nrkcrs encoding small prolinc-rich protciilS. Gellmtlics 16:630- J E: Psoriasin binds calci ulll alt d is upregulated hy calcium ro levels t.h at 637.1993 resemble dI ose observed in normnl skill .) 11I'1('Sf DCI'II/flwl 103:370-375. 1994 3. l3 ackcndorf C . Hohl 0: A CO l11m on origin for cornified envelopc protcins? N nf 23. Saintigny G. Schmidt R . Shroot B, Juhlin L. Reichcrt U . Michel S: Differentia.1 Cellcl 2:91, '1992 cxpression of calgranulin A ,111d B in various cpirhcli al ce ll lines and recon­ 4. Engclkamp 0, Sch ij fcr BW. Mattei MG. Ernc p. l-Ieizmanll CW: Six 5 100 genes structed epidermis.) IIIlIeSI DCl'llIalo/ 99:639-644 . 19')2 arc clustcred 011 human chrom osome 1q2 1: identificOl ti on of two genes coding: 24. Gelb UO . Edclson J G, Desnick RJ: Linkage of p),cnod),sostosis to chromosome for the two previollsly unreported calcium-binding proteins S I 000 Olnd S l ODE. l q21 by homozygosity mapping. NI" C Nwt 10:235-237. 1995. P,..,c Narl Acad Sci USA 90:6547-655 1, 1993 25 . PoJ)' ntcropoulos MH. Ortiz de Luna R.I , Ide SE. Torres R. RubellStein J . 5. Schafer JJW. Wicki R . Engclkamp D . M::lttci M G, H cizmann CW: 1501"tion o f Francomal1 o C A: The gelle fur p),cllud),sosrusis IllapS to humall chro mosome a YAC clone covering a cluster of nine 5100 gcnes on human chromosome Icen-lq21. Nat Cwel IO:238-2.W. 1995 1'1 2 1: r:1tionalc for a new nomenclature of the S I 00 calcium- binding protein 26. Carson S. W il es MV: FOlr upslrealll regions of class II MHC Ea arc necessary for famil y. GCllo",jcs 25:638-643. 1995 position-i.1ldepC ll dcnt. cop),-depclldell t exprcssion of Ea transgclle. N ile/ric 6. Lee S-C, Kim I-G. Marekov LN. O 'Keefe EJ. Parry DAD. Steinert PM : The Acids Res 2 1:2065-2072, I ~93 structure of human trichohya li n: porenti;ll lI1ul tiple fu nctio ns as an EF-hand­ 27. Dillon N. Grosveld F: T r01 ll scriptional regulation of muitigcll e loci: III ult:iI c"cl like cn lciulI1 binding pro tein , a cornified cell cnvelope precursor :lnd an control. TrC'lH/s G W t:I 9: 134 - 137. 1993 intermcdi:ltc fi i:IIl1 Cl1 t nssociated (crosslinking) protein . J Mol Bioi 268:2 164- 2M. Ro,hn"gel J A. Longle y MA. Bundlllan OS. Naylor SL, Lalley PA , J enkins NA. 2 176. 1993 Gilbert DJ. Copeland NG. R oop DR: Charactcrizarion o f ("he mouse loricrin 7. Markova NG. Marekov LN. C hipev CC. Gan 5-Q. Idler WW. Steinert PM : gene: lin kage with proftlaggrin and the Aak y wit and soft coat mutant loci 011 Proftl aggrin is it ll1;ljor epidcnnal c:tl ciull1 binding protein . Ailol Cell Bio/ chromosome 3. Geu(J/llics 23:450-456, 1994 13:(,13-625, 1~93 29. Mosely WS. Seldin MF: Definition OflllOlISC dll'olnosolllc 1 and 3 genc linkage 8. Prcsland R.B. l3 ass uk jA, Kimball JR, Dale BA: C haracterization of two distinCl groups that nrc conserved 0 11 hUIl101n chrol1losmnc I : evidence th:lt ,I conservcd c;llc iul11 - binding sites ill Lite O1 ll1ino-tcrminlls of human protilaggrin. J l/ll /~s l lin kage group spans the centromere of human chromosome I. Gel/olllirs 0,.,.,/111101 104:2 1H-223. 1995 5 : 899-~05. I ~89