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https://tel.archives-ouvertes.fr/tel-00865880
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Étude cristallographique du domaine catalytique de
l’intégrase du virus RAV-1 (rous associated virus type 1)
et découverte d’une nouvelle interface de dimérisation
Allison Ballandras
To cite this version:
Allison Ballandras. Étude cristallographique du domaine catalytique de l’intégrase du virus RAV-1 (rous associated virus type 1) et découverte d’une nouvelle interface de dimérisation. Sciences agricoles. Université Claude Bernard - Lyon I, 2010. Français. �NNT : 2010LYO10251�. �tel-00865880�
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" 38 OKTCU"" <"" Ownvkrng"Kuqoqtrjqwu"Tgrncegogpv"ykvj"Cpqocnqwu"Uecvvgtkpi" ONX" " <" Owtkpg"Ngwmgokc"Xktwu" OO" " <"" ocuug"oqnfiewncktg" Op" " <" ocpicpflug" oqn" " <" oqng" OXX" " <" Ocgfk"Xkupc"Xktwu" OY" " <"" Oqngewnct"ygkijv" P/" " <" Cokpq/" PE" " <" pwenfiqecrukfg" PNU" " <" Pwengct"Nqecnkucvkqp"Ukipcn" RCIG" " <" fingevtqrjqtflug"gp"ign"fg"rqn{cet{ncokfg" Rd" " <" rcktg"fg"dcugu" RET"" " <" Rqn{ogtcug"Ejckp"Tgcevkqp"" RGI" " <" rqn{fivj{nflpg"in{eqn" RFD"" " <" dcpswg"fg"uvtwevwtgu"vtkfkogpukqppgnngu"Å"Rtqvgkp"Fcvc"Dcpm"Ç" rK" " <" rqkpv"kuqfingevtkswg" EKR" " <" eqorngzg"fg"rtfi/kpvfiitcvkqp" Rhw" " <" CFP"rqn{ofitcug"vjgtoqtfiukuvcpvg"kuqnfig" "rctvkt"fg"R{tqeqeewu"hwtkquwu" RRV" " <" rqn{"rwtkpg"vtcev" TCX/3"" <" Tqwu"Cuuqekcvgf"Xktwu"v{rg"3" Tnkdtg"*Thtgg+" <" hcevgwt"fÓceeqtf"T"nkdtg" TOP" " <"" Tfiuqpcpeg"Ocipfivkswg"Pwenficktg" TOUF"" <" Tqqv"Ogcp"Uswctg"Fgxkcvkqp" TPcug" " <" tkdqpwenficug" TUX" " <" Tqwu"Ucteqoc"Xktwu" u" " <"" ugeqpfg"" UCF"" " <" Ukping/ycxgngpivj"Cpqocnqwu"Fkhhtcevkqp" UFU" " <" fqffie{nuwnhcvg"fg"uqfkwo" UKX" " <" Ukokcp"Koowpqfghkekgpe{"Xktwu" UHX" " <" Ukokcp"Hqco{"Xktwu" UKT"" " <" Ukping"Kuqoqtrjqwu"Tgrncegogpv" UKTCU""" <""" Ukping"Kuqoqtrjqwu"Tgrncegogpv"ykvj"Cpqocnqwu"Uecvvgtkpi" Vcs" " <" CFP"rqn{ofitcug"vjgtoqtfiukuvcpvg"fg"Vjgtowu"cswcvkewu" VG" " <" vcorqp"Vtku/GFVC" VGOGF" <" P.P.PÓ.PÓ/vfivtcofivj{n/fivj{nflpgfkcokpg" VH"" " <" vtcpuhqtofig"fg"Hqwtkgt" Vtku" " <" 4/cokpq/4/*j{ftqz{ofivj{n+/3.5"rtqrcpgfkqn" W" " <" wpkvfi" w0c" " <" wpkvfi"cu{ofivtkswg" WK" " <" wpkvfi"kpvgtpcvkqpcng" WX" " <" tc{qppgogpv"wnvtcxkqngv" X" " <"" xqnv"" ¥p" " <"" |kpe" " " " " " " " " " " "
" 39 " " " " " " " " " " " " " " " " " "
Cxcpv/rtqrqu"
" " " " " " " " " " " " " " " " " " " " " " " " " " "" 3: " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " "
" 3; " Eg"vtcxckn"fg"vjflug"c"ffidwvfi"gp"ugrvgodtg"42290"Kn"c"fivfi"tficnkufi"gp"eqvwvgnng"uqwu"nc" fktgevkqp" fw" Rt0" Rcvtkeg" Iqwgv" cw" ugkp" fw" Ncdqtcvqktg" fg" DkqEtkuvcnnqitcrjkg" " nÓKpuvkvwv" fg" Dkqnqikg"gv"Ejkokg"fgu"Rtqvfikpgu"*WOT"72:8+"gv"uqwu"nc"eqfktgevkqp"fw"Ft0"Eqtkppg"Tqphqtv" fcpu" nÓfiswkrg" Tfivtqxktwu" gv" Kpvfiitcvkqp" Tfivtqxktcng" fw" Ncdqtcvqktg" fg" Tfivtqxktwu" gv" Rcvjqnqikgu"Eqorctfigu"*WOT"976+0"
"
" Oqp" rtkpekrcn" uwlgv" fg" tgejgtejg" hwv" nÓfivwfg" etkuvcnnqitcrjkswg" gv" oqnfiewncktg" fw" fqockpg" ecvcn{vkswg" egpvtcn" fg" nÓkpvfiitcug" fw" Tqwu" Cuuqekcvgf" Xktwu" v{rg" 3" *TCX/3" KP" EEF+0" Egu" vtcxcwz" qpv" rgtoku" fg" tfiuqwftg." rct" tgorncegogpv" oqnfiewncktg." nc" uvtwevwtg" fg" TCX/3"KP"EEF" "3.:"¦"fg"tfiuqnwvkqp0"NÓgzcogp"fg"egvvg"uvtwevwtg"tfixflng"swg"ng"fkofltg"fg" nÓKP" fw" TCX/3" rgwv" uÓcuugodngt" uwkxcpv" wpg" pqwxgnng" kpvgthceg" oqnfiewncktg" uvcdknkufig" rct" vtqku" rcktgu" fÓjfinkegu" g0" Egvvg" uvtwevwtg." rct" uqp" cuugodncig" swcvgtpcktg" kpcvvgpfw" gv" ngu" gzrfitkgpegu" fg" ectcevfitkucvkqp" kp" xkvtq" gv" kp" uknkeq" swk" qpv" uwkxk." uwiifltgpv" swg" nÓkpvfiitcug" rquuflfg"fgu"rtqrtkfivfiu"uvtwevwtcngu"oqfwncdngu"gv"qwxtg"fg"pqwxgnngu"rgturgevkxgu"vcpv"uwt"ng" rncp"hqpevkqppgn"swg"ofiecpkuvkswg0"
" Eg" ocpwuetkv" ffietkv" nÓqdvgpvkqp" fg" egvvg" pqwxgnng" uvtwevwtg" gv" ngu" cpcn{ugu" dkqejkokswgu"gv"uvtwevwtcngu"swk"gp"qpv"ffieqwnfigu0" " " Cw"eqwtu"fg"egu"vtqku"cppfigu"fg"vjflug."lÓck"fiicngogpv"gw"nÓqrrqtvwpkvfi"fg"oÓkornkswgt" fcpu"wp"ugeqpf"uwlgv"fg"tgejgtejg"rqtvcpv"uwt"ng"fqockpg"gzvtcegnnwncktg"fg"nc"ufitkpg1vjtfiqpkpg" mkpcug"Uvm3"fg"Uvcrj{nqeqeewu"cwtgwu0"Eg"uwlgv."kuuw"fg"nc"eqnncdqtcvkqp" "nÓKDER"gpvtg"ngu" Rtu0"Dgtvtcpf"Fwenqu"gv"Rcvtkeg"Iqwgv."c"hckv"nÓqdlgv"fÓwpg"vjflug"tficnkufig"gv"uqwvgpwg"rct"ng"Ft0" Rcvtkekc"Rctcewgnnqu"gp"422;0"
Uvm3" c" ffil " fivfi" kfgpvkhkfig" eqoog" tgurqpucdng" fg" nc" rjqurjqt{ncvkqp" fg" rnwukgwtu" gp|{ogu"kornkswfigu"fcpu"ng"ofivcdqnkuog"egpvtcn"fg"nc"dcevfitkg"ckpuk"swg"fcpu"ngu"rjfipqoflpgu" fg"xktwngpeg"gv"fg"tfiukuvcpeg" "nÓcpvkdkqvkswg"rjqurjqo{ekpg0"Ngu"fqockpgu"gzvtcegnnwncktgu" fkvu" Å"RCUVC"Ç" uqpv" urfiekhkswgu" fgu" Ugt1Vjt" mkpcugu" gv" fgu" Rgpkeknnkp/Dkpfkpi" Rtqvgkpu" gv" uqpv"rtqdcdngogpv"kornkswfiu"fcpu"nc"tgeqppckuucpeg"fw"rgrvkfqin{ecpg0"Nc"eqppckuucpeg"fg" nc"uvtwevwtg"fg"egu"fqockpgu"ejg|"Uvm3"rqwttckv"ugtxkt"fg"dcug" "nc"eqpegrvkqp"tcvkqppgnng"fg" pqwxgcwz"kpjkdkvgwtu" "xkufig"vjfitcrgwvkswg0" Pqwu"pqwu"uqoogu"kpvfitguufiu"rnwu"rctvkewnkfltgogpv"cwz"vtqku"fqockpgu"RCUVC"fg"nc" rtqvfikpg"Uvm30"Nqtu"fg"egvvg"fivwfg."cwewpg"ectcevfitkucvkqp"uvtwevwtcng"pÓcxckv"fivfi"eqpfwkvg"uwt" egvvg"mkpcug0"Ckpuk."pqwu"cxqpu"tfiwuukv" "tfiuqwftg"ng"rtqdnflog"fgu"rjcugu"gp"gptgikuvtcpv"fgu" fqppfigu"OCF" "vtqku"nqpiwgwtu"fÓqpfg"*rke."kphngzkqp."jcwvg"fipgtikg+"cw"ugwkn"M"fw"|kpe" " rctvkt"fÓwp"etkuvcn"fkhhtcevcpv"lwuswÓ "4.;"¦"fg"tfiuqnwvkqp0" Eg"ugeqpf"uwlgv"pg"ugtc"rcu"ffixgnqrrfi"fcpu"eg"ocpwuetkv0"Pficpoqkpu."kn"oÓc"crrqtvfi" wpg"xkukqp"rnwu"nctig"fgu"vgejpkswgu"wvknkucdngu"rqwt"tfiuqwftg"wpg"uvtwevwtg"etkuvcnnqitcrjkswg." rnwu" rctvkewnkfltgogpv" ng" rjcucig" rct" ffitkxcvkqp" fÓcvqogu" nqwtfu0" Gp" qwvtg." egvvg" fivwfg" oÓc" rgtoku"fg"oÓkpkvkgt"cwz"vgejpkswgu"fg"fqemkpi"swg"lÓck"rw."gpuwkvg."crrtqhqpfkt"fcpu"ng"vtcxckn" fg"vjflug"rtfiugpvfi0" " " Rwdnkecvkqpu"uwt"nÓkpvfiitcug"<" Dtgxgv"tfihfitgpeg"EPTU"<"GR"324;2738"
Dcnncpftcu." C0." Tqdgtv." Z0." Oqtgcw." M0." Jcugt." T0." Tqphqtv." E0" (" Iqwgv" R0" *4232+0" Crrnkecvkqpu" kpfwuvtkgnngu" gv" ofifkecngu" fÓkpjkdkvgwtu" ekdncpv" nc" pqwxgnng" hqtog" qnkiqofitkswg"fgu"kpvfiitcugu"tfivtqxktcngu0"
" 42 Dcnncpftcu."C0."Oqtgcw."M0."Tqdgtv."Z0."Jcugt."T0."Tqphqtv."E0"("Iqwgv."R0"*4232+0"Vjg"et{uvcn"
uvtwevwtg" qh" vjg" ecvcn{vke" eqtg" fqockp" qh" cp" Cxkcp" Ngwmcgokc" cpf" Ucteqoc" Xktwu" kpvgitcug"tgxgcnu"c"pqxgn"fkogtke"cuuqekcvkqp0"Uqwoku0"
" "
Rwdnkecvkqpu"uwt"nc"Ufitkpg1Vjtfiqpkpg"mkpcug"<"
Rctcewgnnqu." R0." Dcnncpftcu." C0." Tqdgtv." Z0." Eq||qpg." C0L0." Fwenqu." D0" (" Iqwgv." R0" *422;+" Et{uvcnnk|cvkqp"cpf"kpkvkcn"Z/tc{"fkhhtcevkqp"uvwf{"qh"vjg"vjtgg"RCUVC"fqockpu"qh"vjg" Ugt1Vjt" mkpcug" Uvm3" htqo" vjg" jwocp" rcvjqigp" Uvcrj{nqeqeewu" cwtgwu0" Cevc" Et{uv0" Ugev0"H."87."33:9/33:;0"
"
Rctcewgnnqu."R0."Dcnncpftcu."C0."Tqdgtv."Z0."Mcjp."T0."Eq||qpg."C0L0."Fwenqu."D0"("Iqwgv."R0" *4232+0"Vjg"gzvgpfgf"eqphqtocvkqp"qh"vjg"40;"¦"et{uvcn"uvtwevwtg"qh"vjg"vjtgg/RCUVC" fqockp" qh" vjg" Ugt1Vjt" mkpcug" htqo" vjg" jwocp" rcvjqigp" Uvcrj{nqeqeewu" cwtgwu0" L0" Oqn0"Dkqn0."fqk<32032381l0lod042320320234."uqwu"rtguug0" " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " "
" 43 " " " " " " " " " " " " " " " " " "
Gzrqufi"dkdnkqitcrjkswg"
" " " " " " " " " " " " " " " " " " " " " " " " " " "" 44 " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " "
" 45
K0"Tfivtqxktwu"
" Ngu"tfivtqxktwu"crrctvkgppgpv" "nc"hcoknng"fgu"Tgvtqxktkfcg"gv"kphgevgpv"rtkpekrcngogpv" ngu"xgtvfidtfiu0"Egvvg"hcoknng"tgitqwrg"75"gurflegu"kfgpvkhkfigu."tfirctvkgu"gp"fgwz"uqwu/hcoknngu"gv" ugrv"igptgu"encuufiu"ugnqp"ng"Eqokvfi"Kpvgtpcvkqpcn"fg"Vczqpqokg"fgu"Xktwu"*KEVX+"gp"hqpevkqp" fg" ngwt" rcvjqifipkekvfi" gv" fg" nc" uvtwevwtg" fg" ngwt" ifipqog" *Vcdngcw" 3+0" Eg" uqpv" fgu" xktwu" gpxgnqrrfiu" "CTP"oqpqecvfipcktg"fg"rqnctkvfi"rqukvkxg."nÓkphqtocvkqp"ifipfivkswg"fivcpv"rqtvfig" rct" fgwz" oqnfiewngu" fÓCTP" nkfigu" fg" hc›qp" pqp" eqxcngpvg0" Fcpu" ng" xktwu" ocvwtg." nÓCTP" ifipqokswg"guv"cuuqekfi"cwz"rtqvfikpgu"fg"pwenfiqecrukfg"*PE+"gv"nÓgpugodng"guv"gphgtofi"fcpu" wpg" ecrukfg" *EC+" eqpkswg0" Wp" pkxgcw" fÓgpecrukfcvkqp" uwrfitkgwt" eqortgpf" nÓgpxgnqrrg" nkrkfkswg" rtqxgpcpv" fg" nc" egnnwng" kphgevfig." swk" guv" gptkejkg" gp" rtqvfikpgu" xktcngu" *rtqvfikpgu" Å"uwthceg"wpkv"Ç"UW"gv"vtcpuogodtcpcktg"VO+0"Nc"hceg"kpvgtpg"fg"egvvg"gpxgnqrrg"guv"vcrkuufig" rct" nc" rtqvfikpg" fg" ocvtkeg" *OC+0" Nc" vcknng" fw" xktwu" gpvkgt" guv" fÓgpxktqp" 342" pcpqoflvtgu" *Hkiwtg"3+0""
"
Hkiwtg" 3"<" Oqtrjqnqikg" fÔwpg" rctvkewng" tfivtqxktcng0" Ngu" fkhhfitgpvu" eqorqucpvu" uqpv" kpfkswfiu"<" PE"
*pwenfiqecrukfg+."EC"*ecrukfg+."OC"*ocvtkeg+."TV"*tgxgtug"vtcpuetkrvcug+."RT"*rtqvficug+."KP"*kpvfiitcug+."VO" *gpxgnqrrg"vtcpuogodtcpcktg+"gv"UW"*rtqvfikpg"fg"uwthceg+0"
"
"
Ng"e{eng"tfirnkecvkh"fgu"tfivtqxktwu"guv"ectcevfitkufi"rct"nÓfivcrg"fg"vtcpuetkrvkqp"kpxgtug"fg" nÓCTP" xktcn" gp" CFP" dkecvfipcktg0" Eg" fgtpkgt" guv" gpuwkvg" kpvfiitfi" cw" ugkp" fw" ifipqog" fg" nc" egnnwng"j»vg"rct"wpg"rtqvfikpg"xktcng"<"nÓkpvfiitcug0"Fgu"gttgwtu"nqtu"fg"nc"vtcpuetkrvkqp"kpxgtug" uwtxkgppgpv" htfiswgoogpv." kpfwkucpv" ckpuk" wpg" itcpfg" xctkcdknkvfi" ifipfivkswg" cw" ugkp" fgu" tfivtqxktwu0"C"rctvkt"fw"ifipqog"kpvfiitfi."crrgnfi"cwuuk"rtqxktwu."uqpv"vtcpuetkvu"ngu"iflpgu"xktcwz" swk" rgtogvvgpv." crtflu" firkuucig" gv" vtcfwevkqp." nc" hqtocvkqp" gv" nÓcuugodncig" fg" pqwxgnngu" rctvkewngu" kphgevkgwugu0" Ngu" tfivtqxktwu" rgwxgpv" ‒vtg" vtcpuoku" fg" ocpkfltg" gzqiflpg" uwkvg" " nÓkphgevkqp"fg"nc"egnnwng"rct"wpg"rctvkewng"xktcng."qw"fg"ocpkfltg"gpfqiflpg"rct"vtcpuokuukqp"fw" ifipqog"xktcn"kpvfiitfi"fg"nc"egnnwng"ofltg" "nc"egnnwng"hknng0""
" 46
Itqwrg"XK"Î"Tfivtqxktwu" "CTP"ukorng"dtkp" Hcoknng" Uqwu"hcoknngu" Igptgu" Gurflegu"v{rgu"
Tgvtqxktkfcg" Qtvjqtgvtqxktkpcg" Cnrjctgvtqxktwu" Cxkcp"ngwmquku"xktwu" Dgvctgvtqxktwu" Oqwug"ocooct{"vwoqt"xktwu" Icooctgvtqxktwu" Owtkpg"ngwmgokc"xktwu"" Fgnvctgvtqxktwu"" Dqxkpg"ngwmgokc"xktwu"" Gruknqptgvtqxktwu" Ycnng{g"fgtocn"ucteqoc"xktwu" Ngpvkxktwu" Jwocp"koowpqfghkekgpe{"xktwu"3"
Urwoctgvtqxktkpcg" Urwocxktwu" Ukokcp"hqco{"xktwu"
Vcdngcw" 3"<" Vczqpqokg" fgu" tfivtqxktwu" ugnqp" nc" encuukhkecvkqp" fivcdnkg" rct" ng" Eqokvfi" Kpvgtpcvkqpcn" fg"
Vczqpqokg"fgu"Xktwu"*KEVX+"gp"422;0"
"
Fcpu"nc"uqwu/hcoknng"fgu"Qtvjqtgvtqxktkfcg."qp"fkuvkpiwg"<""
3/"Ngu"xktwu"crrctvgpcpv"cwz"igptgu"cnrjctgvtqxktwu."dgvctgvtqxktwu."icooctgvtqxktwu." fgnvctgvtqxktwu" gv" gruknqptgvtqxktwu" swk" qpv" wp" rqvgpvkgn" qpeqifipkswg" rqwxcpv" kpfwktg" fgu" vwogwtu"xctkfigu0"Egtvckpu"rqtvgpv"wp"qpeqiflpg"kpfwkucpv"fgu"vwogwtu"cki¯gu"cxge"wpg"rfitkqfg" fg" ncvgpeg" vtflu" eqwtvg." vgnu" swg" ngu" Cxkcp" Ucteqoc" Xktwugu" *CUX+0" Knu" uqpv" ifipfitcngogpv" ffirqwtxwu" fÓwpg" rctvkg" fw" ifipqog" tfivtqxktcn" gv" uqpv" ffihkekgpvu" rqwt" nc" tfirnkecvkqp0" Egu" qpeqxktwu"uqpv"cnqtu"ceeqorcipfiu"fÓwp"xktwu"eqornfiogpv."vgn"swg"ng"Tqwu"Cuuqekcvgf"Xktwu" *TCX+."rquufifcpv"ngu"vtqku"iflpgu"fg"uvtwevwtg"pfieguucktgu" "nc"tfirnkecvkqp0"FÓcwvtgu"xktwu."vgnu" swg" ngu" Cxkcp" Ngwmgokc" Xktwu" *CNX+." pg" rqtvgpv" rcu" fÓqpeqiflpgu" ocku" uqpv" cevkxcvgwtu" fÓqpeqiflpgu"egnnwncktgu0"Ngwt"ncvgpeg"guv"dgcweqwr"rnwu"nqpiwg0"Egu"fgwz"v{rgu"xktcwz"CUX"gv" CNX" uqpv" tgitqwrfiu" cw" ugkp" fÓwpg" o‒og" gurfleg"<" ngu" xktwu" CUNX" *Cxkcp" Ucteqoc" cpf" Ngwmgokc"Xktwugu+0""
4/" Ngu" ngpvkxktwu" uqpv" pqp" qpeqiflpgu" gv" tgurqpucdngu" fg" ocncfkgu" ejtqpkswgu" " fixqnwvkqp"ngpvg"fqpv"ngu"ukipgu"enkpkswgu"pg"ug"ocpkhguvgpv"swÓcrtflu"fgu"oqku"xqktg"fgu"cppfigu0" Ng"tgrtfiugpvcpv"ng"rnwu"eqppw"guv"ng"xktwu"fg"nÓkoowpqffihkekgpeg"jwockpg"*XKJ+."cigpv"ecwucn" fw"u{pftqog"fÓkoowpqffihkekgpeg"ceswkug"*UKFC+0"
Ngu" urwocxktwu" qpv" wp" oqfg" fg" tfirnkecvkqp" rctvkewnkgt" gv" pg" rtfiugpvgpv" rcu" fg" rcvjqnqikg"cuuqekfig."eg"swk"ngu"rnceg"cw"ecttghqwt"fgu"jfircfpcxktwu."fgu"tfivtqvtcpurquqpu"gv" fgu" tfivtqxktwu" eqorngzgu0" Egu" rctvkewnctkvfiu" qpv" eqpfwkv" " nc" etficvkqp" fÓwp" pqwxgcw" uqwu/ itqwrg" rctok" ngu" tfivtqxktwu." ngu"Urwoctgvtqxktkpcg" *Ngjocpp/Ejg" gv" cn0." 4228+0" Ng" Ukokcp" Hqco{"Xktwu"*UHX+"crrctvkgpv" "eg"igptg0" " "
"30"Ng"ifipqog"tfivtqxktcn"CTP1CFP"
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