Seminars in Cancer Biology 14 (2004) 387396

Viral interactions wit te !otc "atway

S# $iane %aywar&
Viral Oncology Program, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University,
School of edicine, C!" #$%, &'($ Orleans Street, "altimore, ) *&*#&, US+
Abstract
'e !otc signaling "atway in(l)ences cell (ate &ecisions* "roli(eration +ers)s &i((erentiation an& cell s)r+i+al#
Vir)ses ,ot )tili-e an&
mani")late te &i((erentiation state o( in(ecte& cells* "romote or ,loc. cell cycling an& em"loy a +ariety o( mecanisms
to e+a&e innate
cell)lar anti/+iral res"onses an& "romote cell s)r+i+al# 0n ligt o( tese commonalities* it is "era"s not s)r"rising tat
se+eral +ir)ses a+e
ta""e& into te !otc "atway to a&+ance teir own li(e cycles# 'is (irst ,ecame a""arent (rom st)&ies sowing
targeting o( 1"stein/Barr
+ir)s "roteins to te n)clear e((ector o( !otc signaling CS2 (CB3145B67.)# 8ore recently te 9a"osi:s sarcoma/
associate& er"es+ir)s
5'; "rotein as ,een (o)n& to ,in& CS2# !otc "atway interactions a+e also ,een &escri,e& (or a&eno+ir)s SV40
an& )man "a"illoma
+ir)s# 'is re+iew (oc)ses on te er"es+ir)s "rotein interactions wit te !otc "atway an& te insigts tat tese
interactions a+e
"ro+i&e&#
< 2004 1lse+ier 2t&# ;ll rigts reser+e&#
Key,ords- !otc "atway= 1"stein/Barr +ir)s= 9a"osi:s sarcoma er"es+ir)s
1. Introduction
1"stein/Barr Vir)s (1BV) an& 9a"osi:s sarcoma/associate&
er"es+ir)s (9S%V) are )man er"es+ir)ses wit
1BV ,elonging to te gamma 1 lym"ocry"to+ir)s gro)"
an& 9S%V te gamma 2 ra&ino+ir)s gro)"# ; caracteristic
o( er"es+ir)ses is teir a,ility to set )" a latent state &)ring
wic only a s),set o( te +iral genes are e>"resse& an&
te in(ecte& cell s)r+i+es to esta,lis a li(e/long in(ection in
te ost ?1*2@# %er"es+ir)ses also )n&ergo a lytic re"licati+e
"ase in wic te in(ecte& cell &ies an& te +irions "ro&)ce&
s"rea& te in(ection to nai+e cells an& new osts# 2ytic re"lication
ta.es "lace in cells tat are growt arreste& ?3@# 1BV
esta,lises latency in B cells an& it is te latency 1B!;2 an&
1B!;3 "roteins tat interact wit CB31an& "romote B cell
"roli(erati+e res"onses# 1BV can also in(ect e"itelial cells
were te lytic +ers)s latent o)tcome a""ears to ,e more
&elicately ,alance&# 'e latency B;5' enco&e& "roteins
568S an& 59/B;530 are not well/caracteri-e& ,)t teir
+..reviations- 1BV* 1"stein/Barr +ir)s= 9S%V* 9a"osi:s sarcomaassociate&
er"es+ir)s= 1B!;2* 1"stein/Barr +ir)s n)clear antigen 2=
%6V* )man "a"illoma +ir)s= ;&V* a&eno+ir)s= SV40* simian +ir)s 40=
!otc0C* intracell)lar &omain o( !otc= CS2* < "romoter/,in&ing (actor*
S)""ressor o( airless= 2ag/1= S906* S.i/interacting "rotein= 2861*
latency mem,rane "rotein 1= 5';* re"lication an& transcri"tion acti+ator=
B;5'* "am%0/; rigtwar& transcri"ts
'el#A +1/410/9BB/2B48= (a>A +1/410/B02/6802#
/0mail address- &aywar&CDmi#e&) (S#$# %aywar&)#
transcri"ts are igly e>"resse& in e"itelial cells an& tese
"roteins may mo&i(y !otc acti+ity in te e"itelial setting
?46@# 9S%V esta,lises latency in B cells an& also in(ects
en&otelial cells# 0n contrast to 1BV* 9S%V a""ears to rely
"rimarily on mani")lation o( te Ent &e+elo"mental "atway
to "romote "roli(erati+e res"onses ?7*8@# 0t is te 9S%V
lytic cycle reg)lator 5'; tat interacts wit CB31 ?9*10@#
2. EBV EBNA2 interaction with CBF1
1B!;2 is a transcri"tional transacti+ator tat is one o(
te (irst +iral genes e>"resse& a(ter 1BV in(ection o( B cells
an& is essential (or 1BV immortali-ation o( B cells in +itro
?11*12@# 1B!;2 &oes not ,in& $!; &irectly ,)t reF)ires
interactions wit cell "roteins (or "romoter targeting# 5egions
o( te 1BV latency "romoters reF)ire& (or 1B!;2
res"onses were ma""e& an& (o)r gro)"s )sing sligtly &i((erent
strategies i&enti(ie& te 1B!;2/interacting "rotein
as CB3145B67. ?1316@# CB31 (C"/,in&ing (actor 1) is te
)man omolog o( $roso"ila s)"ressor o( airless (S)%)*
m)rine recom,ination/,in&ing "rotein 7 .a""a (5B67_) an&
C1 elegans 2ag/1# 'ese "roteins are o(ten re(erre& to as
CS2 (or CB31* S)% an& 2ag/1# ;t te time o( tis i&enti(ication*
CS2 "roteins were .nown (rom genetic e>"eriments
to ,e &ownstream e((ectors o( !otc signaling# %owe+er*
te mecanisms ,y wic te !otc signal le& to canges
in gene reg)lation in te n)cle)s were not well/)n&erstoo&#
1044/B79G4H see (ront matter < 2004 1lse+ier 2t&# ;ll rigts reser+e&#
&oiA10#10164D#semcancer#2004#04#018
388 S1)1 Hay,ard 2 Seminars in Cancer "iology &3 4*$$35 #%67#8'
3ig# 1# (;) Scematic o( te CB31 core"ressor com"le># 'e locations o( te S906* S85' an& C05/,in&ing sites relati+e to te CB31
re"ression &omain
are in&icate&# 'e same m)tations tat "re+ent S85' ,in&ing to CB31 also "re+ent C05 ,in&ing ?21*22@# (B) Scematic o( te
interactions in te
acti+ation com"le># 'e similarity o( targeting o( !otc0C an& 1B!;2 is igligte&# 0nteractions wit S906 occ)r tro)g te
an.yrin re"eat &omain
(!otc0C) an& Conser+e& 5egion B (C5B* 1B!;2)# 0nteractions wit te re"ression &omain )tili-e te 5;8 (!otc0C) an& C56
(1B!;2) &omains#
'ranscri"tional acti+ation o( target genes occ)rs tro)g te com,ination o( loss o( re"ression an& te "ositi+e e((ects o( te acti+ation
&omain (;C')#
!* amino termin)s= C* car,o>y termin)s#
St)&ies wit 1B!;2 "ro+i&e& some insigt an& re+eale&
tat 1B!;2 mimic.e& !otc0C ()nction to a remar.a,le
&egree#
'ransient assays )sing Ial4/CB31 constr)ctions re+eale&
tat CB31 ()nctione& as a transcri"tional re"ressor# CB31
me&iates re"ression tro)g &irect contacts wit te ,asal
transcri"tion macinery ?17@ an& tro)g cromatin remo&eling#
'e CB31 re"ressor com"le> incl)&es S85'4!Co5*
%$;C1* %$;C2* S;630* C05 an& S906 ?1822@# 0n te
core"ressor com"le>* S906 ,in&s to te car,o>y/terminal
50$/2 &omain o( S85' an& along wit S85' is im"ortant
(or n)clear entry o( CB31 ?21@ (3ig# 1;)# S906 also interacts
wit mSin3; ?19@ an& tis along wit te (act tat S;630
is "art o( te mSin3 com"le> im"lies tat mSin3 is also
"resent in te com"le># 5ecently 80!'4S%;56 as ,een
i&enti(ie& as a CB31 interacting core"ressor wose ,in&ing
is (acilitate& ,y S906 ?23*24@# S%;56 ,in&s S85' ,)t as
also ,een (o)n& to associate wit te !)5$ com"le> ?2B@#
1B!;2 ,in&s to te re"ression &omain o( CB31* con+erting
CB31 into a transcri"tional acti+ator ?26@# 'is con+ersion
is a two/ste" "rocess as &emonstrate& )sing 1B!;2
m)tants# ;n 1B!;2 ca"a,le o( ,in&ing CB31 ,)t lac.ing
te acti+ation &omain was a,le to relie+e re"ression ,y
&is"lacing te CB31core"ressor com"le># 'ranscri"tional
)"reg)lation reF)ire& te "resence o( te 1B!;2 acti+ation
&omain wic recr)its coacti+ator "roteins ?26*27@#
!otc10C an& !otc20C "ro+e& to interact wit te same
&omain o( CB31 an& again tere was e+i&ence (or te o)tcome
,eing te s)m o( relie( o( re"ression "l)s acti+ation
?18*28*29@# 'e contacts ma&e ,y 1B!;2 on te CB31
re"ression &omain a""ear to ,e &is"lace& C/terminally relati+e
to tose ma&e ,y !otc0C an& "oint m)tations in CB31
can &iscriminate ,etween 1B!;2 an& !otc0C/,in&ing#
3or e>am"le* a tri"le alanine s),stit)tion o( 92V 249/2B1
in CB31 a,olises !otc10C an& !otc20C/,in&ing ,)t
&oes not a((ect 1B!;2/,in&ing ?29@ wile s),stit)tions
at 2326 an& J333 &iminis 1B!;2 interaction ,)t not
!otc10C/,in&ing ?30@#
'e mecanism o( !otc0C an& 1B!;2 &is"lacement
o( te CB31 core"ressor com"le> a""ears to ,e i&entical
(3ig# 1B)# 0n eac case a&Dacent &omains ma.e contact wit
CB31 an& S906 to &is"lace S85'# S906 is "artic)larly
interesting as it is a com"onent o( te core"ressor com"le>
an& it is also a com"onent o( te acti+ation com"le>#
$irect com"etition (or ,in&ing to S906 as ,een &emonstrate&
(or S85' an& 1B!;2 an& (or S85' an& !otc0C
?19*20@# Similarly* ,in&ing o( S85' to CB31 is m)t)ally
e>cl)si+e wit ,in&ing to 1B!;2 or !otc0C ?18*19@# 'e
5;8 &omain o( !otc0C is te maDor contact wit CB31
?28*29*31@ wile te an.yrin re"eats ,in& S906 ?20@# Conser+e&
5egion B o( 1B!;2 ,in&s S906 an& Conser+e& 5egion
6 ,in&s CB31# ; m)tation witin Conser+e& 5egion 6
tat a,olises 1B!;2 ,in&ing to CB31 (EE323S5) com"letely
a,olises 1B!;2 transacti+ation ?32*33@ an&* wen
incor"orate& into te +ir)s* m)tation o( tese two resi&)es
res)lts in a non/immortali-ing 1BV ?34@# 8)tants in Conser+e&
5egion B s)c as 00307 tat are im"aire& (or S906
interaction are eF)ally im"aire& (or transacti+ation ()nction
?19@# 0ntro&)ction o( &eletions tat s"an C5B o( 1B!;2 into
te 1BV genome le& to m)tant +ir)s tat eiter (aile& to immortali-e
B cells in +itro or res)lte& in B cell colonies tat
grew less well tan wil&/ty"e +ir)s immortali-e& controls
S1)1 Hay,ard 2 Seminars in Cancer "iology &3 4*$$35 #%67#8' 389
?3B@# 0nterestingly* it was note& in te latter case tat te cells
o+ere>"resse& te m)tant 1B!;2 "rotein# ; similar sit)ation
is (o)n& wit !otc10C were m)tation o( two amino
aci&s in te (o)rt an.yrin re"eat a,olise& S906 interaction*
se+erely im"aire& transacti+ation o( a re"orter containing
CB31/,in&ing sites ?20@ an& a,olise& te a,ility o( !otc
to ini,it myogenesis ?36@# ')s* 1B!;2 an& !otc0C eac
reF)ire contacts wit S906 as well as CB31 (or e((ecti+e
acti+ation o( "romoters containing CB31/,in&ing sites#
*1&1 od9lation of /":+* transcriptional responses
'ere are ()rter "arallels ,etween 1B!;2 an& !otc0C
in te ways in wic transcri"tional acti+ation tro)g
CB31 is mo&i(ie&# 'e acti+ation &omains o( tranacti+ator
"roteins ,in& coacti+ators tat mo&i(y cromatin str)ct)re
tro)g istone acetylase acti+ity an& ma.e contacts wit
te ,asal transcri"tional macinery# 'e acti+ation &omain
o( 1B!;2 ,in&s core transcri"tional (actors ?37*38@* "300*
6C;3 an& CB6 ?27@ an& a coacti+ator* "100 tat interacts
wit '3001 ?39@# 0n a&&ition transacti+ation is a)gmente&
,y ,in&ing to te 1BV enco&e& 1B!;/26 "rotein ?40*41@
an& ,y interactions wit te SE04S!3 com"le> ?42@ an&
te $1;$/,o> "rotein $61034Iemin3 ?43@# 'e !otc0C
acti+ation &omain ,in&s IC!4* 6C;3 an& "300 ?4446@
an& ()ll transcri"tional acti+ation ,y !otc reF)ires te )man
omolog o( $roso"ila 8astermin&* 8;821 wic
,in&s to te an.yrin re"eat &omain o( !otc an& also recr)its
CB64"300 ?4749@# 5es"onses to !otc an& 1B!;2
are igly concentration/&e"en&ent an& negati+e reg)lation
o( te acti+ity o( te CB31 com"le> occ)rs tro)g mo&)lation
o( CB31 $!; ,in&ing# 1BV enco&es tree relate&
"roteins generate& tro)g gene &)"lication* 1B!;3;* 3B
an& 3C# 'ese "roteins com"ete wit 1B!; 2 (or ,in&ing
to CB31 ?B0B3@# 'e CB311B!;3 com"le> &oes not
,in& to $!; ?B4*BB@ an& so transcri"tional acti+ation is
,loc.e& ,ot ,y loss o( access to CB31 an& ,y loss o( CB31
"romoter targeting# 'e "ositi+e reg)lation o( 1B!;2 ,y
1B!;/26 ,in&ing an& te ,alancing negati+e reg)lation
,y te 1B!;3 (amily "laces 1B!;2 acti+ity largely )n&er
+iral control an& at te le+el o( te CB31 com"le> (3ig# 2)#
!otc0C acti+ity is also reg)late& in "art ,y loss o( CB31
,in&ing# 'e 208 "rotein 9yo'2 was i&enti(ie& in a yeast
two/y,ri& screen as a CB31/interacting "rotein an& sown
to "re+ent $!; ,in&ing ,y CB31 ?B6@# 80!'4S%;56
antagoni-es !otc0C ,in&ing to CB31 ?23*24@# K+erall*
owe+er* !otc0C ()nction may ,e reg)late& less at te
"oint o( CB31 com"le> an& more tro)g "rocesses s)c as
&i((erential e>"ression o( !otc ligan&s* rece"tor recycling
an& "rotein t)rno+er ?B7*B8@# 'e mecanistic concor&ance
,etween te interactions o( 1B!;2 an& !otc0C wit te
CB31 re"ressor com"le> is re(lecte& in te a,ility o( te
two "roteins to acti+ate te same CB31/reg)late& genes
in re"orter assays ?B9@* 1BV latency "romoters (wit te
e>ce"tion o( 2861) ?60@ an& a((ect e>"ression o( te same
cell)lar genes# 1>am"les o( te latter are )"reg)lation o(
3ig# 2# 0ll)stration o( 1BV mo&)lation o( te CB31 com"le># CB31 ,in&s
to te seF)ence I'III;; ?143*144@# 1B!;2 )ses CB31 (or "romoter
targeting an& mimicry o( !otc signaling# 'e 1B!;3 (amily o( "roteins
negati+ely reg)late 1B!;2 acti+ity ,y ,in&ing to CB31 an& "re+enting
CB31 (rom ,in&ing $!; ?B4*BB@ an& ,y com"eting wit 1B!;2
(or CB31/,in&ing ?B0*B3@# 568S "re+ents 1B!;2 &is"lacement o( te
core"ressor com"le> ?4@# 1B!;/26 interaction wit 1B!;2 increases
1B!;2 acti+ity ?40*41@#
B;'3 ?61@ an& C$21 ?60@ an& &ownreg)lation o( te 0g
enancer ?60*62@#
!otc0C me&iates e((ects tat are in&e"en&ent o( CB31
?636B@ an& 1B!;2 can target "romoters tro)g 6)#1
?66*67@# 1B!;2 an& !otc a+e no "rimary seF)ence omology
an& eac a+e s"eci(ic "rotein "artners# %owe+er*
CB31 targeting a""ears to ,e a &ominant ()nction (or ,ot
"roteins to te e>tent tat !otc0C an& 1B!;2 can ,e
"artially e>cange& in some ()nctional assays# !otc0C
"re+ents te &i((erentiation o( C2C12 myo,lasts into myot),)les
an& 1B!;2 as a similar e((ect ?68@# 0n tose cells
con&itionally e>"ressing 1B!;2* !otc10C co)l& "artially
com"ensate (or loss o( 1B!;2# 0n one st)&y* !otc10C
s)""orte& cell "roli(eration (or a sort "erio& in te "resence
o( an in&e"en&ently e>"resse& 2861 gene ,)t tere
was a &e(icit in )"reg)lation o( c/8yc an& te cells &ie&
a(ter se+eral &ays ?69@# 0n a secon& st)&y* iger le+els o(
!otc10C resc)e& cell growt alto)g te cells e>"an&e&
more slowly tan in te "resence o( 1B!;2 ?70@# 2ym"o,lastoi&
cell lines are &e"en&ent on 2861 (or growt
an& te ina,ility o( !otc0C to e((ecti+ely )"reg)late 2861
is consistent wit te 2861 "romoter a+ing more com"le>
reg)lation tan sim"le CB31 tetering# 1B!;2 also ,in&s
to a 6)#1 site in te 2861 "romoter ?66*67@* an& )niF)ely*
1B!;3 co/o"erates wit 1B!;2 rater tan antagoni-ing
1B!;2 acti+ation o( 2861 e>"ression ?71*72@#
*1*1 /":+* and cell s9rvival
0n lym"o,lastoi& B cell lines e>"ressing 1B!;2 ()se&
to te ormone ,in&ing &omain o( te estrogen rece"tor*
wit&rawal o( estrogen res)lts in cessation o( cell growt
an& a signi(icant "ro"ortion o( te cells &ie ,y a"o"tosis
?73@# 2oss o( e>"ression o( 1B!;2/reg)late& genes s)c as
cell)lar c/myc* cyclin $2 an& c&.4 ?73*74@ an& 1BV 2861
an& 2862 ('6) ?7B@ contri,)te s),stantially to tis o)tcome#
2861* wic ()nctionally mimics constit)ti+e t)mor
necrosis (actor rece"tor signaling* "romotes cell s)r+i+al
390 S1)1 Hay,ard 2 Seminars in Cancer "iology &3 4*$$35 #%67#8'
tro)g acti+ation o( !3/_B* Bcl/2 an& ;20 ?7683@ an&
;.t acti+ity ?84@# 2862; "ro+i&es a cell s)r+i+al ()nction
also "artially tro)g acti+ation o( ;.t ?8B87@# $ata
o,taine& )sing emo"oietic "rogenitor cells (rom !otc
antisense transgenic mice in&icates tat !otc1 can reg)late
!3/_B acti+ity ?88@ an& e+i&ence as ,een "resente&
(or !otc10C/in&)ce& )"reg)lation o( Bcl/2 an& 60394;.t
acti+ity in ' cells ?89@# ; com"onent o( te co/o"erati+e
e((ect o( !otc1 on trans(ormation ,y )man "a"illoma
+ir)s (%6V) 16 an& 17 is anti/a"o"totic an& me&iate&
tro)g ;.t ?90@# !otc10C also as te a,ility to "rotect
against 'C5/in&)ce& a"o"tosis tro)g a &irect interaction
wit te n)clear ormone rece"tor (amily mem,er !)r77
?91@# !)r77 acts ,ot as a transcri"tion (actor ?92@ an&
as a me&iator o( a"o"tosis# 0n te latter ca"acity* !)r77
translocates to te cyto"lasm an& targets mitocon&ria to
in&)ce cytocrome C release ?93@# 'e anti/!)r77 acti+ity
o( !otc10C "rom"te& an e+al)ation o( te anti/a"o"totic
ca"a,ilities o( 1B!;2# Lsing Sin&,is +ir)s in(ection to
in&)ce !)r77 syntesis an& a"o"tosis* it was &emonstrate&
tat 1B!;2 co)l& li.ewise ,in& to !)r77 an& "rotect
against !)r77/in&)ce& cell &eat ?94@# 'e mecanism
o( "rotection was i&entical# Bin&ing o( eiter !otc10C
or 1B!;2 "re+ente& mitocon&rial targeting o( !)r77 in
res"onse to a"o"totic stim)li an& !)r77 remaine& in te
n)cle)s#
*1#1 Cell cycle arrest- /":+* and KSHV !T+
!otc me&iates cell (ate &ecisions an& can "romote &i((erentiation
as well as "roli(eration# 0n "rimary .eratinocytes
!otc10C ca)ses growt s)""ression tro)g in&)ction o(
"21 ?9B@ an& a similar o,ser+ation as ,een ma&e a(ter
!otc o+ere>"ression in small cell l)ng cancer cells ?96@#
!otc10C e>"ression in te cic.en B cell line $'40 also
in&)ces I1 cell cycle arrest ?97@# 'e &i((erentiation res"onse
to !otc can ,e tiss)e/s"eci(ic or relate& to a com,ination
o( te &e+elo"mental stage o( te cell an& te "resence
or a,sence o( mo&)lators o( signal intensity# Eile
1B!;2 is clearly growt "roli(erati+e in te conte>t o( an
1BV in(ection* e>"ression o( 1B!;2 in te a,sence o( te
mo&)lating 1B!;3 "roteins res)lts in retar&ation o( cell
growt an& "21 in&)ction# 1B!;2 e>"ression in naso"aryngeal
carcinoma cells* osteosarcma L2KS* Vero an& 293
cells le& to "21 in&)ction me&iate& tro)g "B3 an& retar&ation
o( cell growt ?98@# B)r.itt lym"oma B cells maintain
ty"e 0 latency an& &o not e>"ress 1B!;2 or te 1B!;3
"roteins# Con&itional e>"ression o( 1B!;2 in B)r.itt lym"oma
cells res)lte& in growt arrest an& in&)ce& en&ogeno)s
inter(eron al"a ?99*100@# 1>ogeno)s e>"ression o(
1B!;2 in ;.ata B)r.itt lym"oma cells in&)ce& te 1BV
lytic cycle* wic ta.es "lace in I1 arreste& cells* an& tis
in&)ction reF)ire& te CB31 interacting &omain o( 1B!;2
?101*102@# 0n 1BV "ositi+e lym"o,lastoi& cells con&itional
o+ere>"ression o( 1B!;3; li.ewise ca)se& I04I1 growt
arrest ?B3@# ')s* e+en in B cells* te res"onse to 1B!;2 is
igly &e"en&ent on te le+el o( "rotein a+aila,le (or CB31
interaction#
9S%V in(ects ,ot B cells an& en&otelial cells# 'e
9S%V 5'; "rotein is te .ey +iral "rotein reg)lating te
switc (rom latent to lytic +iral re"lication ?103*104@# 5';
is a transcri"tional transacti+ator tat ,in&s &irectly to $!;
,)t also targets +iral "romoters in&irectly ,y tetering to
cell)lar transcri"tion (actors ?10B*106@# Kne o( te cell)lar
"artners (or 5'; is CB31 ?9@# 'e regions o( CB31 necessary
(or 5'; ,in&ing incl)&e te re"ression &omain tat is
also targete& ,y 1B!;2 an& !otc0C# ')s* 5'; a""ears to
,e )sing te same mecanism to acti+ate CB31 containing
"romoters# ; &i((erence ,etween 1BV an& 9S%V in teir
mimicry o( !otc is tat 1BV* tro)g 1B!;2* )ses te
"atway (or +iral latent in(ection wile te 9S%V 5'; "rotein
is te .ey reg)lator o( te lytic cycle an& 9S%V latent
in(ection can ,e esta,lise& in CS2 n)ll cells ?10@# 'e &i((erent
)se o( te !otc "atway may ,e relate& in "art to te
cell tro"ism o( 9S%V# !otc ()nction is critical to +asc)lar
&e+elo"ment ?107*108@ an& in en&otelial cells !otc signaling
"romotes cell &i((erentiation ?109@# 'e lytic re"licati+e
"ase o( a er"es+ir)s li(e cycle ta.es "lace in cells tat
are arreste& in te cell cycle an& 9S%V enco&es lytic "roteins
tat (acilitate cell cycle arrest ?110@# 'o &ate 5'; as
only ,een sown to acti+ate 9S%V genes tro)g CB31 ,)t
i( 5'; also targets CB31/reg)late& en&otelial cell genes*
te o)tcome so)l& ,e com"ati,le wit a lytic re"licati+e
en+ironment (or te +ir)s#
3. EBNA2 function in EBV biology
1BV immortali-es B cells in c)lt)re to generate contin)ally
"roli(erating B lym"o,lastoi& cell lines# 1B!;2 an&
te 1B!;3; an& 3C "roteins are essential (or tis "rocess
as is 2861 ?111@# 1B!;2 clearly stim)lates B cell s)r+i+al
an& growt "roli(eration in te setting o( 1BV in(ection#
3)rter* as alrea&y esta,lise&* 1B!;2:s main mecanism
o( action is as a mimic o( !otc0C# 0t as not ,een clear ow
to integrate tese (acts wit wor. &emonstrating tat a &ominant
()nction o( !otc10C in emato"oiesis is to in&)ce
common lym"oi& "rogenitors to a&o"t a ' cell (ate at te
e>"ense o( B cell &e+elo"ment ?11211B@# ;lto)g te role
o( !otc in te B cell com"artment remains incom"letely
)n&erstoo&* recent o,ser+ations allow s"ec)lation on te
ways in wic te 1B!;2 )s)r"ing o( !otc ()nction may
s)stain a li(e/long 1BV in(ection in te ost# 3irstly* tere
is e+i&ence tat !otc signaling increases te (ormation
o( emato"oietic stem cells# 7agge&1 is e>"resse& on ,one
marrow stoma an& on stromal cell lines an& coc)lt)re o(
m)rine marrow "rec)rsors wit 7agge&1 increase& te (ormation
o( "rec)rsor cell "o")lations ?116*117@# Ksteo,lastic
cells also e>"ress 7agge&1 an& increase& e>"os)re to osteo,lasts
a)gments "rimati+e aemato"oietic cell growt
in a !otc/&e"en&ent manner ?118@# 1B!;2/&ri+en B cell
"roli(eration may t)s ,e reca"it)lating a "rec)rsor stem
S1)1 Hay,ard 2 Seminars in Cancer "iology &3 4*$$35 #%67#8' 391
cell res"onse# 'is "roli(eration is to)gt to ,e im"ortant
in e>"an&ing te in(ecte& cell "o")lation a(ter "rimary 1BV
in(ection to "ermit esta,lisment o( a li(e/long latency in
te B cell com"artment#
0n ealty 1BV/"ositi+e in&i+i&)als* e>"ression o(
1B!;2 is restricte& to nai+e 0g$+ B cells in secon&ary
lym"oi& tiss)e ?119*120@# 0g$+ B cells are concentrate&
in te marginal -one o( germinal centers# 'ransgenic mice
tat are con&itionally &e(icient (or CB31 in B cells sow
a selecti+e loss o( marginal -one B cells wit loss o(
CB31* im"licating !otc signaling in te &e+elo"ment o(
tese cells ?121@# 5ein(orcing tis o,ser+ation* trans(er o(
80!'/&e(icient s"lenic B cells into reci"ient mice le& to
"re(erential &i((erentiation into marginal -one B cells ?24@#
80!'4S%;56* a CB31 interactor tat antagoni-es !otc
acti+ity* as ig e>"ression in mo)se (ollic)lar B cells an&
low e>"ression in marginal -one B cells# !otc2 is te rele+ant
!otc (amily mem,er as !otc2 e>"ression is ig in
marginal -one B cells ?122@ an& ()rter* con&itional &eletion
o( !otc2 in mice res)lte& in a &e(iciency in marginal
-one B cells ?123@# 'e (act tat 1B!;2 is selecti+ely
e>"resse& in te same com"artment o( te germinal center
were !otc2 e>"ression is essential (or B cell &e+elo"ment
strengtens te conce"t tat 1BV is )sing 1B!;2 as
a +irally controlle&* constit)ti+e !otc s)rrogate#
. !he EBV BA"!s
'e 1BV Bam%0/; rigtwar& transcri"ts (B;5's) were
(irst &escri,e& in naso"aryngeal t)mor tiss)es were tey
are te most a,)n&ant +iral transcri"ts ?124*12B@# B;5's
were s),seF)ently recogni-e& to ,e e>"resse& in all 1BV
latently in(ecte& cells ?126130@# 'e alternati+ely s"lice&
B;5' (amily o( 5!;s contain tree recogni-e& o"en rea&ing
(rames# 0t as ,een &i((ic)lt to con+incingly &emonstrate
e>"ression o( te "roteins enco&e& ,y tese 5!;s in
1BV/in(ecte& cells ,)t te "ro&)cts o( two o( te o"en rea&ing
(rames* 568S an& 59/B;53K interact wit te !otc
"atway wen e>"resse& e>ogeno)sly# 568S ()nctions in
a way tat is reminiscent o( 80!'4S%;56# 568S ,in&s to
C05 in te CB31 core"ressor com"le> an& me&iates re"ression
wen e>"resse& as a Ial4/568S ()sion "rotein ?4@#
568S antagoni-es 1B!;2 ()nction in re"orter assays an&
!otc10C ()nction in ,ot re"orter assays an& in a m)scle
&i((erentiation assay ?4*B@# 568S a((ects te C05S906 interaction
in a way tat "re+ents 1B!;24!otc10C (rom &is"lacing
te core"ressor com"le># S906 (acilitates S%;56
()nction an& t)s S906 is not only a .ey tetering "oint (or
1B!;24!otc0C ,in&ing ,)t also a .ey target (or negati+e
mo&)lation o( teir acti+ity#
'e 1BV 59/B;530 "rotein was (o)n& to interact wit
!otc4 in a yeast two/y,ri& assay ?6@# 59/B;530 contains
a "otential signal "e"ti&e (or en&o"lasmic retic)l)m targeting
an& interacts wit )n"rocesse& !otc4# 59/B;530 a""ears
to "otentiate !otc ()nction# 'e B;5's are e>"resse&
in circ)mstances in wic te 1B!;2 an& 1B!;3 "roteins
are not syntesi-e& s)c as in e"itelial cell in(ection an& in
1BV/associate& t)mors in imm)nocom"etent in&i+i&)als# 0t
is interesting tat tese transcri"ts so)l& enco&e ,ot a "ositi+e
an& a negati+e !otc reg)lator an& tis re/em"asi-es
te im"ortance o( signal mo&)lation (or !otc "atway res"onses#
'e e>istence o( te B;5's also s)ggests tat tere
may ,e as yet )na""reciate& as"ects o( 1BV ,iology were
mani")lation o( !otc signaling occ)rs#
#. Adeno$irus% hu&an 'a'illo&a $irus and (V)
Kter +ir)ses tat a+e interactions wit te !otc
"atway are a&eno+ir)s (;&V)* %6V an& simian +ir)s 40
(SV40)# 'e (irst association wit ;&V came (rom te
recognition o( a CB31(5B62!) ,in&ing site in te "0G
"romoter ?131@# S),seF)ently ;&V 13S11; was sown to
,in& to CB31 an& to ,e ca"a,le o( acti+ating "romoters
containing CB31/,in&ing sites in re"orter assays ?132@#
13S11; interaction reF)ire& CB31 seF)ences !/terminal
to te CB31 re"ression &omain ,)t te mecanism o( action
still a""ears to in+ol+e &is"lacement o( te S85'
core"ressor com"le>#
'e interactions ,etween !otc an& %6V an& !otc an&
SV40 are co/o"erati+e in nat)re# !otc1 ,)t not !otc2 can
&ownreg)late e>"ression o( te %6V 16417 genes in cer+ical
cancer cells# 'e &ownreg)lation res)lts (rom s)""ression
o( ;6/1 acti+ity an& increase& e>"ression o( te negati+e
reg)lator 3ra1 ?133@# %6V16 16 an& 17 "roteins in t)rn )"reg)late
!otc1 acti+ity tro)g increase& transcri"tion an&
tro)g increase& e>"ression o( "resenilin/1 ?134@# 'ese
o,ser+ations s)ggest te e>istence o( a (ee&/,ac. loo" to
reg)late !otc acti+ity# 'e co/o"erati+ity ,etween !otc0C
an& 16417 in trans(ormation assays as ,een s)ggeste& to
&eri+e (rom an anti/a"o"totic ()nction me&iate& tro)g ;.t
?90*13B@# SV40 in(ection also transcri"tionally )"reg)lates
e>"ression o( !otc1 ?136@# SV40 in(ection as ,een associate&
wit )man mesotelioma ?137@# 0n )man mesotelial
cells* te SV40/me&iate& increase in !otc1 acti+ity
is necessary (or cell growt as treatment o( te cells wit
a gamma/secretase ini,itor le& to I248 cell cycle arrest
?136@#
*. Conclusion
'e a,ility o( !otc signaling to in(l)ence ,ot "roli(erati+e
an& &i((erentiation res"onses ma.es tis "atway
an attracti+e target (or +ir)ses wic are sensiti+e to te
&i((erentiation state o( te cell# %6V an& SV40 ta" into
te "atway ,y )"reg)lating e>"ression o( acti+ate& !otc
wile 1BV* 9S%V an& ;&V enco&e "roteins tat ,in& to
CB31 an& mimic as"ects o( !otc signaling# 1ac o( tese
+ir)ses also as mecanisms to &ysreg)late cell cycle control#
;cti+ate& !otc signaling in a cell in wic cell cycle
392 S1)1 Hay,ard 2 Seminars in Cancer "iology &3 4*$$35 #%67#8'
cec."oints a+e ,een ren&ere& ine((ecti+e esta,lises a
s)sce"ti,ility to )nreg)late& growt# %6V* 1BV an& 9S%V
are associate& wit )man cancers an& SV40 as ,een lin.e&
to mesotelioma# 0n te cases o( %6V an& SV40/associate&
cancers were te +iral "roteins )"reg)late !otc acti+ity*
&r)gs &irecte& against !otc ()nction or "rocessing
may a+e treatment +al)e i( a tera"e)tic win&ow e>ists in
wic normal !otc signaling in te "atient can ,e s"are&#
9S%V targets CB31 &)ring te +iral lytic cycle# ;lto)g
lytically 9S%V/in(ecte& cells are ,elie+e& to contri,)te to
9S%V/associate& t)morigenesis tro)g "aracrine signaling*
tere are oter "atways acti+ate& in te lytic cycle tat
wo)l& seem to "ro+i&e more attracti+e anti/+iral targets#
1BV as in+este& ea+ily in mani")lation o( !otc signaling
wit 1B!;2* 1B!;3;* 3B an& 3C* 1B!;/26* 59/B;530
an& 568S all ,eing &irecte& towar&s te CB31 com"le>#
'ese "roteins are e>"resse& in 1BV/associate& t)mors in
imm)nocom"romise& "atients ?138141@# ; cell/"ermea,le
1B!;2 "e"ti&e tat ,loc.s te 1B!;2CB31 interaction
can "re+ent 1BV immortali-ation o( "rimary B cells in +itro
?142@ "ro+i&ing "roo( o( "rinci"le tat targeting te !otc
"atway may a+e clinical "otential against 1BV/associate&
&isease# 'ese 1B!;s are also transcri,e& in secon&ary
lym"oi& tiss)e in ealty sero"ositi+e in&i+i&)als ?119@
an& it is interesting to consi&er weter s)ccess()l targeting
o( tis "atway co)l& eliminate li(e/long 1BV in(ection#
Ac+nowledge&ents
'is wor. was s)""orte& in "art ,y grant 537 C;4224B
(rom te !ational Cancer 0nstit)te#