3. Primary antibodies
!"#$% &'()%"*+%"*),-'+.%)./$% 0'12,*3%+,45),'*6 07%
0'12,*3%+,45),'*6
8&9% 9'#/"*.:('51;$%
Anti-A.*+ /ouse monoc3ona3 *56++ A7cam
Anti-.3a8 9a77it po3yc3ona3 *56++ *5*+++ ;i8ma
Anti-<A=> ?@A-A? /ouse monoc3ona3 *5>+++ A? Aioscience
Anti-<A=> ?@A-A? /ouse monoc3ona3 *5>+++ A? Aioscience
Anti-8eminin (.=-2+D) 9a77it po3yc3ona3 *56++-*5*+++ ;anta-CruH
Anti-8eminin 9a77it po3yc3ona3 *56++-*5*+++ =in8Iei =uo
Anti-<.P /ouse monoc3ona3 *5*+++ Chemicon
Anti-<.P /ouse monoc3ona3 *5*+++ /oAiLec
Anti-<.P /ouse monoc3ona3 *5*+++ 9oche
Anti-<;L /ouse monoc3ona3 *5*++++ @oMa8en
Anti-NA (O.*+) 9at monoc3ona3 *56++ *526+ 9oche
Anti-@P/ /ouse monoc3ona3 *56++ *52++ A7cam
Anti-LAP (O<O) /ouse monoc3ona3 *56+++ IrQin ?aMi%son
Anti-LAP /ouse monoc3ona3 *56+++ /artin Leichmann
Anti-LAP=* (2A*) /ouse monoc3ona3 *56++ *5*++ IrQin ?aMi%son
Anti-L.IIA /ouse monoc3ona3 *5O+++ /artin Leichmann
Anti-LIPL 9a77it po3yc3ona3 *56++-*5*+++ *5*++-*5*+++ /ara Pitu3escu
Anti-!-tu7u3in /ouse monoc3ona3 *5>+++ ;i8ma
Anti-"-tu7u3in (<LR-88) /ouse monoc3ona3 *5*++-*52++ A7cam
Table 10. Lhe primary anti7o%ies use% in this stu%y.
Abbreviations
111
A""#$%&'(&)*+
AA. acrylamide-bisacrylamide 3AT. 3-amino-17 27 4-tria:ole ActD. actinomycin D
AdE4. adenovirus E4 promoter
AdMLP. adenovirus maBor late promoter APS. ammonium persulphate
ATM. ataxia telangectasia mutated bHLH. basic helix-loop-helix bp. base pair
BRE. TFIIB binding element BREd . BRE doLnstream BREu . BRE upstream C. cytoplasmic fraction
ChIP. chromatin immunoprecipitation CHO. cycloheximide
CKII. casein kinase II cpm. counts per minute
DAPI. 476-diamidino-2-phenylindol DEPC. diethyl pyrocarbonate DIG. digoxigenin
DMEM. DulbeccoTs modified EagleTs medium DMSO. dimethylsulfoxide
DPE. doLnstream promoter element dpp. days post-partum
DRB. 576-dichloro-1-beta-D-ribofuranosylben:imida:ole EDTA. ethylenediaminetetraacetic acid
EGFP. enhanced green fluorescence protein FACS. Fluorescence-activated cell sorting FCS. fetal calf serum
GFP. green fluorescent protein Gmnn. geminin
!bbreviations
11.
/S13: glycogen synthase kinase 3 /ST: glutathione sulfur transferase HRP: horseradish peroBidase
HSC T1: herpes simpleB virus thymidine kinase IFR: initiator element
IPT/: isopropylthio-!-H-galactoside kHa: kilo Haltons
LB: Luria-Bertani medium Mk: marker
F: nuclear fraction FF1: neurofibromin 1 Fgn.: neurogenin . FPM: nucleophosmin M.H.: optical density oligo: oligonucleotides MRF: open reading frame P: pellet
PBS: phosphate buffered saline PC: Polycomb
p.c.: post coitum Pc/: Polycomb group
PCR: polymerase chain reaction PCC: packed cell volume PF!: paraformaldehyde
PH/PB: phospholipids hydroperoBide glutathione peroBidase PMSF: phenylmethylsulfonyl fluoride
Pol I: RF! polymerase I Pol II: RF! polymerase II Pol III: RF! polymerase III pp: post partum
PRC1: polycomb repressive compleB 1 PRC1L: polycomb repressive compleB 1-like Prm: protamines
!""r$vi'tion+
--.
/012 3o+t$rior +$4 co6"+
78'+$ !2 ri"on9cl$'+$ ! r362 ro9n; 3$r 6in9t$
r78!2 ri"o+o6'l 78!
7<2 roo6 t$63$r't9r$
7<=/172 r$v$r+$ tr'n+cri3tion=3ol>6$r'+$ c?'in r$'ction 02 +93$rn't'nt
0!@A0/@2 +t$ril$ 'l3?' 6otiB ;o6'in 0c6?-2 0$4 co6" on 6i;l$C ?o6oloC - 0D02 +o;i96 ;o;$c>l +9lB't$
+i78!2 +6'll int$rB$r$nc$ 78!
0EI2 +$l$ctivit> B'ctor I
07G2 +93r$++or+ oB 78! 3ol>6$r'+$ G 69t'tion+
0HI2 +9"v$ntric9l'r Jon$
0>t I2 +>n'3tot'C6in I
<!K+2 <G/='++oci't$; 3rot$in+
<GE2 <ri+="or't$=ED<!
<G/2 <!<! "o4 "in;inC 3rot$in
<G/E-2 <!<! "in;inC 3rot$in=liM$ B'ctor
<E@ED2 8N8N8ON8O=t$tr'6$t?>l=$t?>l$n$;i'6in$
<KII2 <r'n+cri3tion B'ctor+ II
<I/<2 <!<! "in;inC 3rot$in=liM$ B'ctor=int$r'ctinC 3rot$in
<62 6$ltinC t$63$r't9r$
<3n2 tr'n+ition 3rot$in+
<7K2 <G/=r$l't$; B'ctor+
t<!K+2 t$+ti+ +3$ciBic <!K+
<9P-2 n$9ron'l t9"9lin Q2 9nit+
QGK2 93+tr$'6 "in;inC B'ctor Q1E2 93+tr$'6 control $l$6$nt H2 volt+
HI2 v$ntric9l'r Jon$
A""re%iations
11.
Ackno&ledge+ents
110
Acknowledgements
"n the 'ir*t +,-.e " /o1,d ,i3e to e4+re** 56 5o*t *in.ere th-n3* to 56 *1+er7i*or Pro'. :r.
;i.h-e, <e**e,= 'ir*t o' -,, 'or gi7ing 5e the .h-n.e to do 56 Ph: in hi* ,-?or-tor6. " -5 re-,,6 gr-te'1, 'or -,, the +-tien.e -nd the ti5e he in7e*ted in o+ening 56 e6e* on /h-t it 5e-n* the re-, *.ien.e. " o/n hi5 - ,ot o' gr-tit1de 'or te-.hing 5e - ne/ /-6 to thin3. "
/o1,d ne7er 'orget hi* -d7i.e*= /hi.h /i,, 'o,,o/ 5e .on*t-nt,6 /here7er the '1t1re dri7e* 5e.
" -5 gr-te'1, to Pro'. :r. @rn*t Ai55er 'or -..e+ting to ?e 56 re'eree -nd -,*o to -,, the 5e5?er* o' 56 Ph: .o55ittee= Pro'. :r. B-,' Ci.ner= Pro'. :r. Bo?ert Dr-d*tein= Pro'. :r.
EFrg GtH,3e -nd to Pro'. :r. BHdiger I-rde,-nd.
" /-nt to th-n3 to 56 e4.e,,ent .o,,-?or-tor* :r. Jhri*tine ;-6er= Pro'. :r. "ngrid Dr155t=
Pro'. :r. ;-rtin Tei.h5-nn -nd :r. Beinh-rd <,e5ent 'or their gre-t he,+ -nd -,/-6*
+re.io1* -d7i.e* .on.erning 56 /or3.
;6 *+e.i-, th-n3* to Pri7. :oL. :r. An-*t-**i- Gto63o7- 'or ,ightening 56 ne1ron-, .onne.tion* -nd *1++orting 5e .on*t-nt,6. " /-nt to -ddre** - N;er.i ?e-1.o1+O to the Cren.h *+e-3er* 'ro5 56 de+-rt5ent= Pro'. :r. Ah5ed ;-n*o1ri -nd :r. P-tri.3 Jo,,o5?-t 'or -,, their *1++ort -nd en.o1r-ge5ent* d1ring the di''i.1,t ti5e*. A* the6 -,/-6* to,d 5e=
NA,,e* /ird D1tPO. " /o1,d ,i3e to th-n3 to :r. <-5-, Jho/dh1r6= :r. E-n G.hindeh1tte= :r.
Gi,7io BiLLo,i= :r. I-n* :ieter G.h5itt= Tr-n Jong T1o. 'or intere*ting di*.1**ion* -nd
*1gge*tion*.
To the he-d o' the ;-** G+e.tro5etr6 :e+-rt5ent= :r. Ienning Qr,-1?= -nd to Q/e P,eR5-nn -nd ;oni3- B--?e= " th-n3 'or the 5-n6 +rotein *eS1en.ing= /hi.h .on*tit1ted 7-,1-?,e in'or5-tion 'or 56 /or3. Be*ide* " /o1,d ,i3e to e4+re** gr-tit1de to Pro'. :r.
Ao,'g-ng @nge, -nd Pro'. :r. <-ri5 U-6erni- 'or their .on*ider-?,e he,+. " -5 gr-te'1, to Pro'. :r. :et,e' :oene.3e -nd :r. Ui.o,e I-++e, 'or i5+ort-nt he,+ 'or CAJG -n-,6*i*. "
/o1,d ,i3e to -.3no/,edge :r. V,g- -nd :r. Deni- ;-3-ro7 'or g1iding 56 *te+* into
?io.he5i*tr6 'ie,d -nd 'or their 'riend*hi+.
" h-7e - *+e.i-, th-n3 to Gh-ri' ;-h*1r -nd Petr- B1* 'or their te.hni.-, he,+ -nd h-++6 -t5o*+here .re-ted in the ,-?. " -5 7er6 th-n3'1, to 56 -.t1-, -nd 'or5er ,-? 5-te*= :r.
Acknowledgements
116
!ing%ei !uo* Dr. !ars 0ittler* Dr. 3yun56in 7im* Dr. Dere9 :pieler* :<en Pilars9i* >aisana
?sli* @<onne Aerlings and 0ieC9e Behrens %or sharing good and Cad times. F also Gant to than9 them %or nice discussions* especially to my %riend >aisana %or the good Io9es. Besides F Gould li9e to eJpress gratitude to all the memCers o% the Kruss department %or the %riendly atmosphere and their continous help during the past %eG years. F Gill miss you allL
F am indeCted to DMK-Kraduierten9olleg OPole9ulare Keneti9 der QntGic9lungO %or gi<ing me the opportunity to Ce a memCer o% a scienti%ic community and %or the generously %inancial support. F Gould li9e to eJpress my special than9s to Pro%. Dr. Thomas Pieler* Pro%. Dr. Qrnst 0immer and Panuela Pana%as %or all the seminars and Gor9shops organiSed.
Mor pro<iding eJperimental material F Gould li9e to ac9noGledge Pro%. Dr. Partin Teichmann* Pro%. Dr. FrGin Da<idson* Dr. 6ayhong Chong* Pro%. Dr. Pary UsCorn %or their generosities.
Pany Garm than9s to the Vomanian %riends %rom KWttingen* especially to Q<a and Toni 7reiter* ?leJandra ?ndrei* Pihaela Diaconu* Dana and U<idiu Foan* Xlad and Vodica Pena*
:il<io ViSSoli %or the nice time Ge shared together and their support.
:pecial than9s to Pro%. Dr. ?ndrei ?nghel* Gho alloGed me to Ioin the Biochemistry team in Vomania and to Dr. U<idiu :irCu %or gi<ing me the impulse to %olloG science. ?lso F Gant to than9 to the all memCers o% the Biochemistry Department %rom APM Timisoara %or the nice time Ge had together.
Mrom all my heart* a truelly OThan9 youLY to my parents* Kheorghe-:a<a and Tudorita Panescu* %or their lo<e* sacri%ice and permanent support. F also Gant to than9 to my grandmother Pandita %or her precious help during my years o% study and %or all the prayers. F ha<e special than9s %ull o% regrets to my grandmother Para* my OPamiY* Ghich passed aGay Ce%ore time* %or all the care and lo<e. To the Cest sister one can e<er ha<e* Pihaela* and to my soul mate* Parian* F than9 you deeply %or Ceing neJt to me* alGays. Than9 you Parian %or Ceing my Cest %riend all o<er the years* and %or your unconditional lo<e.
Pii de multumiri celor din tara care au %acut posiCil acest <is* Pica* Diana* Draga Pisco<*
Xictor si Parcela Xanturici. Xa multumescL
:cumpii mei parinti* <a multumesc pentru tot ceea ce ati %acut pentru mineL