Function of Smad: Smad 2 found on the chromosome 18q21 and it is present near the Smad 4 locus in the genome

of humans. The mutation studies identified that 6% of the colon cancer is caused due to the missense mutation either in the MHC or MHC of the Smad 2. !" the process of immunostaining it is identified that #$% of human s%in SCCs sho& the reduce production of this protein or loss of tumor tissues. The loss of Smad2 protein is higher in the poorl" differentiated SCCs. The Smad2 does not directl" 'inds &ith the ()* molecule 'ut it comple+es &ith 'oth Smad, and Smad4. Thus 'inding of snail to the S!- leads to the recruitment of Smad4 through the 'inding of snail &ith Smad,. This process in turn leads to the contri'ution of -MT. The Smad2 deficient s%in is more suscepti'le for the formation of s%in tumors and for the con.ersion of malignant one. The loss of Smad 2 &ill up regulate the acti.ities of Smad, and 4 and 'inds &ith the S!- of snail and H/0. Hence it leads in the increased production of snail and H/0 is said to 'e induced '" Smad2 deletion induced tumorogenesis. Smad 3 is in the chromosome of 11q21 2 22 chromosomes in human. t is frequent in the human colon cancer and in 'reast cancers. Ho&e.er the Smad, production is not lost in s%in SCCs &here as the increased e+pression of it is e+pressed in 'reast cancer. T/0 2 'eta o.er e+pression and follo&ed inflammation is induced '" T3* contri'utes greatl" to&ards the tumor de.elopment. Studies indicate that the Smad, has its o&n tumor suppression function through Smad, mediated T/0 2 'eta function. t is said as the ma4or mediator for T/0 2 'eta5 Smad, ma" either acts as a tumor suppressor or tumor promoter in conte+t dependent manner. Smad 4 is identified &ith the tumor suppressor function in pancreatic cancers. )o& it is characteri6ed as the ma4or mediator of T/0 2 'eta signaling. /erm line mutation of Smad4 induces 4u.enile pol"posis s"ndrome. 7oss of hetero6"gosit" had reported in man" tumors. *nal"sis indicates that the loss of Smad4 has 'een associated in the inacti.ation of 3T-) and 321 5it promotes cell proliferation and it inhi'its apoptosis in cells and it is interrelated &ith T/0 2 'eta induced inflammation hence it accelerates tumor de.elopment and the proliferation of the same.

General mechanism of signal transduction: /0 2 'eta signaling controls most of the cellular e.ents li%e regulation of the em'r"onic process5 immunological5 tumor regulation and &ound healing5 etc. /08'eta is functioned '" the 'inding of the ligand molecules to the cell mem'rane receptor5 thus it acti.ates the c"toplasmic mediators in to the nucleus and thus finall" it regulates the e+pression of their target gene. The ligand is three forms and as T/08'eta 15 25 ,. The cell surface receptors are as T/08'eta 9 and 92. :suall" the Smad signaling from the c"toplasm to the nucleus can performed '" three Smad isomers of the famil" and it can 'e said as Smad 25 ,5 4. !inding of the ligand &ith T/08'eta 92 8 915 phosphor"lation of Smad2 and Smad,5 it 'inds &ith Smad4 and forms trimeric comple+5 translocated to nucleus. This comple+ 'inds &ith S!- and regulates the T/08'eta response genes either directl" or '" action of co factors to the target gene. T/0 2 'eta ligand initiates the 'inding and 'ringing 'oth t"pe and t"pe serine ;threonine %inase receptor to the surface of the cell. t is follo&ed '" phosphor"late the domain of receptor and follo&ed '" the signal transduction '" the phosphor"lation process of Smad proteins. Thus the acti.ated Smad comple+ in turn translocated to the nucleus in association &ith other fe& nuclear co factors and thus it regulates the target gene for its function. The other Smads li%e Smad5 Smad65 Smad# &ill negati.el" regulates T/0 2 'eta signaling path&a" '" competing &ith 98 Smads or '" the action of co Smads it targets the receptor for the purpose of destruction. T/0 2 'eta c"to%ine is a dimmer and is 'eing strengthened '" the h"dropho'ic interactions and further it can 'e strengthened '" ha.ing inter su' unit disulfide 'ridges in most of the cases. The t"pe receptor ha.e characteristic S/S/S/ sequence &here as t"pe phosphor"lation of its /S domain '" the t"pe receptor don<t ha.e this. t is termed as /S domain. The acti.ation of t"pe receptor gets in.ol.ed in the receptor. Hence the acti.e signaling molecule or comple+ is said to ha.e 'oth of its receptors 'ound to the ligand. The primary article I had chosen is TGF Beta Signal Transduction by ! "assague Annu. Rev. Biochem. 1998. 67:75391 "echanism for the receptor acti#ation: This re.ie& had done ma4or accomplishment in the studies related to this signaling. t helps in the elucidation of general mechanisms of T/0 2 !eta signaling and its related factors that in turn help in the acti.ation of the signaling process from the cell surface to ()* or the target gene as the successful one. This is 'etter illustrated in the diagram gi.en in page 1. 2

The T/0 'eta in turn 'rings t&o t"pes of %inases li%e seine;threonine %inases and thus it regulates gene e+pression. =ne of this %inase &ill in turn phosphor"late SM*( proteins5 the" &ill mo.e in to the nucleus and generates transcriptional comple+ of specific ()* 'inding a'ilit". This T/0 famil" came to notice &hen it &as studied in the cloning of acti.in receptor >Mathe&s et al., 1??1@. This *CT 9 >Cheifet6 et al., 1?8#@. !ased on the structural features T/0 2 !eta ha.e t&o receptors as > 11 %da@ and >#$ %da@&ith core pol"peptides of 1$$ 2 1#$ amino acids that includes the signal sequence >Mathe&s et al., 1??15Ten et al., 1??,57in et al.51??2 A *ttisano et al., 1??2@ The spacing of c"steine .aries from one receptor to the other. Ser 21, is present in the receptor gets phosphor"lated '" '" the receptor %inase in a ligand independent manner5 most commonl" that is used for the signaling path&a" >7uo et al., 1?#$@. The ser 161 of receptor gets phosphor"lated '" the receptor the ligand dependent manner >Soucheln"ts%"i et al., 1??6@. The unique feature is of /S domain5 t"pe receptor ha.e this highl" conser.ed 2$ amino acids immediatel" preceding the domain of protein %inase. This region has the speciali6ed sequence as S/S/S/ >Brana et al., 1??4@. The ligand induced phosphor"lation of serine and threonine in the sequence of TTS/S/S/ of receptor '" receptor is required for the acti.ation of signaling. This /S region ser.es as the most important region or the regulator" domain for the signaling of T/0 2 'eta path&a". The 0C!312 inhi'its T/0 2 'eta '" 'inding to the unphosphor"lated /S region of the t"pe phosphor"lated T/0 2 'eta 9 7igand 9eceptor nteractionsD The T/0 2 'eta dimmers are held together '" h"dropho'ic interactions and in most of the cases it also held '" an inter disulfide 'ond. -ach monomer consists of three disulfide 'onds interloc%ed and this structure is %no&n as c"steine %nots >Sun et al., 1??1@. $egulation of the receptor acti#ation: The accessing of T/0 2 'eta ligand &ith its receptor is said to control '" t&o molecules &ith opposing functions. The" can 'e said as proregion of T/0 2 'eta precursor and decorin 'inds to the free T/0 2 'eta. The resolution of the cr"stal structure of )oggin 2 !M3# comple+ &as re.ealed and ho& one of these factors gets 'ind &ith the target and thus the a.oidance of 3 receptor molecules >Huse et al., 1???@. The 'inds effecti.el" &ith the Smad2 in .itro and it enhanced the has similar properties &ith the T/0 2 !eta receptor

phosphor"lation of C 2 serine residues of Smad2 residues.

contact &ith mem'rane receptors. )oggin is a class of decorin5 it inhi'its !M3# and hence it 'loc%s the surface that is required for the interaction of t"pe and t"pe et al., 2$$2@. Smad phosphorylation and acti#ation: The C terminal 1$ residues &hich includes the characteristic SSES motif > serine 461 and 46# required for Smad 2 and Smad 4 comple+ formation@ at the e+treme end of C terminus and it is said as completel" fle+i'le and disordered in 9 Smads > Smad 25,54 comple+@. Hence the phosphor"lation of 9 Smad is said to ta%e place at t&o serine residues in C terminal &ithin the fle+i'le SSES motif >*'dollah et al., 1??#@. Thus the phospor"lation leads to the desta'ili6ation of Smad interaction &ith the S*9* and thus allo&s the dissociation of Smad from the comple+5 further the su'sequent e+posure to the nuclear import region of the Smad MH2 >Eu et al., 2$$$@. The associated proteins said as 9 Smad and Smad4 helps in the nucleation and assem'l" of transcriptional region comple+. The 9 Smad &ill get 'ind &ith Smad4. n the structure of phosphor"lated Smad2 the 'inding &ill ta%es place at p Ser 2 E p Ser motif. 3rimaril" the 'inding is coordinated '" four residues and the" are present in the Smad45 it ser.es as 'ind &ith the p Ser 2 E p Ser motif of the 9 Smads in the heterodimeric Smad comple+ >Bu et al., 2$$1 '@. The nuclear translocation of the receptor acti.ated SM*(s occur and follo& agonist induced phosphor"lation and gets associated &ith SM*( 4. The nuclear translocation of SM*(1 and 2 does not require SM*(4 and it is determined '" the SM*(4 defecti.e cells. Hence the receptor acti.ated SM*(s 'inds &ith SM*(4 and transfers it in to the nucleus >7iu et al., 1??#@. SM*(4 is also getting translocated in to the nucleus in response to T/0 >)a%o et al., 1??#@.Smads ma" directl" get associated &ith the 'asal transcription machiner". The MHC domains of the 9 2 Smads and Smad4 gets interacted &ith transcriptional co factors li%e C!35 3,$$5 *9C1$1 A Smif > !ai et al., 2$$25 (er"nc% et al., 1??85 Cato et al., 2$$25 Massague and Batton 2$$$@.This 9 Smads &ill act as )7S for Smad 1 and Smad ,. This acti.it" is entirel" depends on the phosphor"lation of Smad motifs. Thus the ne+t step of import to the nucleus is studied in relation to the MHC 'inds &ith importin 'eta and it is not &it importin alpha >Curasi%i et al., 2$$1@. (ephosphor"lation is carried out '" unidentified proteins5 >9andall et al., 2$$2@ :'iqutination and proteasome mediated degradation carried out '" the termination of Smad signaling. The Smurf1 targets the Smad1 and Smad1 the destruction in the c"toplasm of unstimulated cells >Fhu et al., 1???@. 4 !M3 receptors >/roppe

%onclusion:

learnt from the signaling process made me clear a'out the signaling

process is 'ased on the succession of the protein 2 protein and protein 2 ()* interactions. t gi.es an idea for understanding the other t&o re.ie&s. t learnt me most of the 'asics that need to concentrate. The structural components that mediates the signaling path&a" gi.es an idea a'out the detailed stud" of those elements enforced in this path&a". This helps me in gaining the %no&ledge of either gaining or loss of this T/0 'eta underlies .arious sorts of de.elopmental disorders such as cancer and fe& other ailments in humans. This signaling path&a" e+plains ho& it is &or%ing and the comple+it" of the path&a". t clearl" states that more &or% &ill readil" gi.e more surprises in this stud" of signaling path&a". Ho&e.er this stud" indicates that the ma4or milestones are &aiting in this research. & GGGGGGGGGGGGGGGGGGG PP R Smad with SARA . TGF receptor, Ligand, co receptor complex, receptor, R Smad with SARA Form a complex ' %o receptor comple(

PP GGGGGGGGGGGGGGGGGGGGGGG *' complex 'ormed with * Smad in tead o' R Smad will !e de tro+ed !+ Pho phata e

R Smad and ligand relea ed R Smad relea e !+ pho phata e and !ind with SARA till the reaction

"o Smad R Smad Ligand complex !ind with "o Smad "o Acti$ator or "o Repre or

R Smad # Ligand # "o Smad # "o Acti$ator # %&A !inding 'actor !ind with %&A molec(le and th( the re)(ired molec(le get prod(ced 5

%&A !inding Factor

Illustration: o represents the T/0 'eta 'inding site of the receptorH first rectangular 'o+ as 9eceptor and the second as T/0 'eta H the T/0 'eta ligand 'inds in the o position and gets 'ounds &ith T/0 'eta receptor comple+. )e+t in the o site co receptor &ill 'inds on &ith S*9* and comple+ formed. 9 Smad &ith 7igand 9eleased. !inds &ith Co Smad and forms 9 Smad5 7igand and Co Smad comple+. !inding of co acti.ators or repressor and ()* 'inding factor forms a comple+ and 'inds &ith ()* molecule and gets transcri'ed '" desired protein production. 9 Smad released from the comple+ and 'inds &ith the first comple+ formed for the repetition of the ne+t c"cle of the same. The un&anted are said to 'e dephosphor"lated '" 3hosphatase. Thus the signaling reaction continues till the need is satisfied. $eference: 1. *'dollah5 S.5 Macias8Sil.a5 M.5 Tsu%a6a%i5 T.5 Ha"ashi5 H.5 *ttisano5 .7.5 and Brana5 I.7. 1??#@. T 'eta 9 phosphor"lation of Smad2 on Ser461 and Ser46# is required for Smad28Smad4 comple+ formation and signaling. I. !iol. Chem. 2725 2#6#822#681. 2. *ttisano 75 Brana I75 Cheifet6 S5 Massague I. 1??2. Cell 68D?#21$8 ,. !ai5 9.J.5 Coester5 C.5 =u"ang5 T.5 Hahn5 S.*.5 Hammerschmidt5 M.5 3eschel5 C.5 and (u"ster5 I. >2$$2@. SM 05 a Smad48interacting acti.ators of T/08'eta signalling. )at. Cell !iol. 45 18121?$. 4. Cheifet6 S5 Beather'ee I*5 Tsang M78S5 *nderson IC5 Mole I-5 et al., 1?8#. Cell 48D4$?211 1. (er"nc%5 9.5 Fhang5 J.5 and 0eng5 E.H. >1??8@. SmadsD transcriptional acti.ators of T/08 'eta responses. Cell 955 #,#2#4$. 6. /roppe5 I.5 /reen&ald5 I.5 Biater5 -.5 9odrigue687eon5 I.5 -conomides5 *.).5 C&iat%o&s%i5 B.5 *ffolter5 M.5 Kale5 B.B.5 !elmonte5 I. C.5 and Choe5 S. >2$$2@. Structural 'asis of !M3 signalling inhi'ition '" the c"stine %not protein )oggin. )ature 4205 6,62642 #. Huse5 M.5 Chen5 J.8/.5 Massague5 I.5 and Curi"an5 I. >1???@. Cr"stal structure of the c"toplasmic domain of the t"pe T/08 'eta receptor in comple+ &ith 0C!312. Cell 965 42124,6.

8. Cato5 J.5 Ha'as5 9.5 Catsu"ama5 J.5 )aar5 *.M.5 and He5 E. >2$$2@. * component of the *9C;Mediator comple+ required for T/0L'eta )odal signalling. )ature 4185 6412646. ?. 7iu 05 3ouponnot C5 MassaguMe I. 1??#. Genes Dev. 11D,11#26# 1$. 7uo CE5 7odish H0. 1??#. EMBO J.16D1?#$281 11. Massague5 I.5 and Botton5 (. >2$$$@. Transcriptional control '" the T/08 !etaL;Smad signaling s"stem. -M!= I. 195 1#4121#14 12. Mathe&s 7S5 Kale BB. 1??1. Cell 61D ?#,282 1,. )a%ao *5 mamura T5 Soucheln"ts%"i S5 Ca&a'ata M5 shisa%i *5 et al. 1??#. EMBO J. 16D1,1,262 14. 9andall5 9.*.5 /ermain5 S.5 nman5 /.S.5 !ates5 3.*.5 and Hill5 C.S. >2$$2@. (ifferent Smad2 partners 'ind a common h"dropho'ic poc%et in Smad2 .ia a defined proline8rich motif. -M!= I. 215 1412116. 11. Soucheln"ts%"i S5 ten (i4%e 35 Mi"a6ono C5 Heldin CH. 1??6. EMBO J. 11D62,12 4$ 16. Sun 3(5 (a.ies (. 1??1. Annu. Rev. Bi !"#s.Bi $ l. %t&u't. 24D26?2?1 1#. Ten (i4%e 35 chi4o H5 0ran6Men 35 Schul6 35 Saras I5 et al. 1??,. On' (ene 8D28#?2 8# 18. Brana I75 *ttisano 75Bieser 95Kentura 05 MassaguMe I. 1??4. )atu&e ,#$D,4124# 1?. Bu5 I.8B.5 Hu5 M.5 Chai5 I.5 Seoane5 I.5 Huse5 M.5 7i5 C.5 9igotti5 (.I.5 C"in5 S.5 Muir5 T.B.5 0airman5 9.5 et al. >2$$1'@. Cr"stal structure of a phosphor"lated Smad2D recognition of phosphoserine '" the MH2 domain and insights on Smad function in T/08'eta signaling. Mol. Cell 85 12##2128?. 2$. Eu5 7.5 Chen5 J.8/.5 and Massague 5 I. >2$$$@. Smad2 nuclear import functions mas%ed '" S*9* and unmas%ed '" T/08 'eta dependent phosphor"lation. )at. Cell !iol. 25 11?2162. 21. Fhu5 H.5 Ca.sa%5 3.5 *'dollah5 S.5 Brana5 I.7.5 and Thomsen5 /.H. >1???@. * SM*( u'iquitin ligase targets the!M3 path&a" and affects em'r"onic pattern formation. )ature 4005 68#26?,.