NCU01005 1 k b NCU07896 NCU07897 1 k 8:A6 Y63 8: 2010. 1. 29.¢  DSDSDS S D m - 2 D m - 1 D...

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Transcript of NCU01005 1 k b NCU07896 NCU07897 1 k 8:A6 Y63 8: 2010. 1. 29.¢  DSDSDS S D m - 2 D m - 1 D...

  • D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

    im -2

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    D D DS S S W

    T ∆d

    im -2

    ∆d mm

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    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

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    ∆d mm

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    D D DS S S W

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    D D DS S S W

    T ∆d

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    8:F10 NCU00344

    8:A6 NCU07897NCU07896

    Ψ63 NCU01005 1 kb

    1 kb

    1 kb

    1

    2 1.6

    3 5

    0.5

    10

    1

    2 1.6

    3 5

    0.5

    10

    1

    2 1.6

    3 5

    0.5

    10

    Honda_Supplementary Fig. 1

  • 8G3

    NCU03486NCU03488NCU03489 NCU03487 NCU03485 366 aa486 aa43 aa397 aa1016 aa

    1 kb

    1

    2 1.6

    3

    10 5

    0.5

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d im

    -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

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    -1

    D D DS S S W

    T ∆d

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    D D DS S S W

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    D D DS S S W

    T ∆d

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    ∆d mm

    -1

    D D DS S S W

    T ∆d

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    -1

    D D DS S S W

    T ∆d

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    ∆d mm

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    D D DS S S W

    T ∆d

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    ∆d mm

    -1

    D D DS S S W

    T ∆d

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    ∆d mm

    -1

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    kb

    A G A C A T A G A A A A C G G G G A T A T A C T G T A T T G T A C A C A T T G C T

    W T

    ∆d m

    m -1

    G C C T T G C G A T G C G A T G C A T T A C C T T G G C C G C A A T G C A C G

    A

    B

    Honda_Supplementary Fig. 2

  • Aspergillus clavatus

    Aspergillus fumigatus

    Aspergillus nidulans

    Aspergillus niger

    Aspergillus terreus

    Chaetomium globosum

    Gibberella zeae

    Magnaporthe grisea

    Neosartorya fischeri

    Phaeosphaeria nodorum

    Neurospora crassa JmjC c c 14581

    JmjC c c 7631

    757 JmjC c c1 JmjC c c 7201

    JmjC c c 7551 JmjC c c 7531

    JmjC c c 13511

    JmjC c c 12691

    JmjC c c 7551

    JmjC c c 13451

    JmjC c c 13651

    JmjC c c1 653Aspergillus oryzae

    JmjC c c1 1073Botryotinia fuckeliana

    JmjC JmjC domain c Cysteine-rich region AT-hook motif

    A

    B DMM-1(CysR2) KG-DVYRCHVCQHKDYTYKLAFCSNPGCSEAYCYGVLYRAFDL-MPQEVLQNERWECPRCLK DNMT3L(CysR) DGYQSY-CSICCSGET---LLICGNPDCTRCYCFECVDSLVGPGTSGKVHAMSNWVCYLCLP .* : * * :* : * :*.**.*:..**: : .. . :* ..* * **

    Honda_Supplementary Fig. 4

  • R1060A R1150A

    ∆3x AT-hook

    DMM-1 JmjC c c 14581

    JmjC JmjC domain c Cys-rich region AT-hook motif

    R1090A

    W T

    ∆d mm

    -1

    +R 10

    60 A

    +R 10

    90 A

    +R 11

    50 A

    +R 10

    60 A &

    R 10

    90 A

    +R 10

    60 A &

    R 11

    50 A

    +R 10

    90 A &

    R 11

    50 A

    +3 x m

    uta tio

    n

    +∆ 3x

    AT -ho

    ok

    digestion: Hpa II, probe: 8:G3

    ∆d im

    -2

    A

    B

    +W T

    ∆dmm-1

    1

    2

    1.6

    3

    5

    0.5

    kb

    10

    Honda_Supplementary Fig. 5

  • 8:G3probe:

    1

    2 1.6

    3 5

    0.5

    kb 10

    D D DS S SDSD D DS S S W

    T ∆d

    im -2

    +P dm

    m- 1: :dm

    m- 1H

    21 6A -H

    AT -F

    LA G

    ∆d mm

    -1

    +P dm

    m- 1: :dm

    m- 1-H

    AT -F

    LA G

    +P ac

    tin ::d

    mm -1-

    HA T-F

    LA G

    +P ac

    tin ::d

    mm -1

    H2 16

    A -H AT

    -F LA

    G

    Honda_Supplementary Fig. 6

  • MPAASHPQAK FDPIPPDLDL HALVEKTPNF HWVLRVSAAQ IRNIGPQEFE RLVYLHVIRG GKPLVIEKWN DRLPKTLFSA EWLEGTYNKK QENVRDIVAQ TDIPMTMGHY LRSMKQLTNQ WTPSNFRDER RQRLYLKDID CPPEWHERLQ KIIPTNLFYM NDNLDKKVSR RFDDYDYVEE KSCAPAGDLM SSLPEEMRAQ NLMCYVGHEG TYTPAHREMC ASLGQNIMVD ASGDENGEKP GSSIWFMTET KDREVVREYF LSILGHDIEI EKHFAQINAW KKANFPVYIV EQKVGDFVLV PPLAPHQVWN RGTRTIKVAW NRTTAETLEM ALHEALPKAR LVCRDEQYKN KAIIYFTLQK YYNQMIEMEQ NADIGLLGFG QDLMKNSFRM TQIAKDFKAL FRLFTEILVD EMFATKEKEI EYIEFDSCIT CSYCRANIFN RFLTCKHCVR TLVTGDEDTY DICMECYAMG RSCLCVSNLT WAEQWKWSDL VDRYEEWRAL IIKNDGYVDY DTSPLPLELA RRKSGKKSVA QICQEQLRRR PWKDISKLDK PSEPENSEPE VDENGKPKKK PRRKKKKGDV YRCHVCQHKD YTYKLAFCSN PGCSEAYCYG VLYRAFDLMP QEVLQNERWE CPRCLKICNC GACRRLKNGN VPYHPKNTLL GHDTRPIADD RSVETLVDFK VHNLNWLKTS GDDGRNVQSK RLQKLQQQAE AEKAKNADVH IQVPALPSGE QATGAENLDD ALQQSIADGL GEDQERPDVE MTGASADQPN VVQPAGPAEV ADMSVAPEDH DQSAYPDPSM FGQERMLGMG YYQQDDDSPD KILFDPYQRP TAENYVVDEE PELMSEYLKK QLRLAKRRAR HAEDDDPDFA APRSHHRKKP KLDKTSQPPP TRDGQDDALE NMDPALFDQT MTDVGGDIRD GSAQAGASQP ANAQSADEAA EESRQARMYS PNVPTLRHAR PKVSYRVDED NEEEFNDILI PRSQRPEYRA AAAPASVVQS IEGPTGPKDP LDLASAAVLA ITGAGGNRSN EASQEPGSAR AAAGSTGPSK LRGPHKKRGR PPGRPRKSEA HHTEDEESHE APEPTRRRGR PPRASLAAKE PADSSHKSGE ENDDEDIDAQ LAQELDGFDA DGEVVERDAG VPQAPKRRGR PPKNGVKKVV VEVTTKAPML SLAERMKLKG KKFKIGERKS TGGTPGKVIA KPVDSVEKSA SPAGSGRVAR DAEKQTSLPV RESPRMTRTR TTSATRHTVQ PAEEESHHSE SEPEAEPEKE MENKKSPEPP ARRSASQASV RISPPRDEAS PNKSPERSPS VSASPEPPNR APSTAAAPSA TRISPSPSPS PVRSPSPPPP PPKPAGPTVV RLMDTEDEES DYGGSYSGSR SPSQRSESER SGSVASGTGS EDEPSSDSDS DDEDIPARKE SVRQSFASRG RGGGVGVVSR GRGGIRGRGR GRGRPRGS

    DMM-1, 162,990.6 Da (1458 aa) 66 unique peptides, 638/1458 amino acids (44% coverage)

    MEALLGASNS NPSCGSILEL REQLRSVFST KVTEKRAEHV SVTLELRSTS VFHIPVTEPE NEQLEQAASI DPSLGGAQSS VAPASGEGGA NGNVNGNQPT RQINAIDALI NQPSDDHVLQ KSVAKHIITT LGEIDGSSWA VRQVSRGEQG WTFGYICKDS WQAWSRQNAK NPGRAVIGEW SQKPDSMDAV NLIGQLIELL VPPPAPREPK HALQRREPKK PKEPKPPKPP KPPKQPKPPK DPNAPKPPRK KRPAGEGGAG DGTQPKKRRK KKDAAAVGPD GSILPPEMPG GDTQNGYSTY PEGLVGQAPA AADEGVHVHS ILNIPPAEAA RRRTVAIDLL ESGGIDPKTL SPEQFNIFAN QAPELQQESL DMLKRYGAER LRIVHPDKEQ GDGAQAAPAT QAATAPSTAP INPLKKKSKK KKSGVVDSEE GVAASGGKKK KKKNKEKLTR GKCDPCRDRK IPCTKEKPSC SQCTTSGLQC HYALERKVKD ISPESEAEPE AQSEPVVVEA DEEPEGLGSP GFHNEPAHHE PASHEPFNQE QDSHDSIDNN MPHGLPTSGL VSPQPESATT YHQPSDLYQQ FNSSVDASAT PEVAHTGVTP AAMDYMHSSV TDNSLHNFSY TQPQSEPEPA SAPVPEPEPI PEPVPVVEQP PPVQHTSNYA QQSSSTQAGG RTRRSLPSGP SVQSGAMNGH TTHAPTESWT PVPVPTIPAA YQNKPSPRAS RARKPAPAAP QTSMQNMTQH RTQNTVQQPA SQAFDDLRQA SSWSSVPQPT IPAQKTSQQQ ARTSPYQAAA QAVRASSRQG NRSQNHTPVQ QQTSAPGRHS QLQASQSHTD NSNYQSNTGV AASTSVGNYD SYSQYPTSSS RTEATNTRMN YEPYTNPSST TSNSYSSYDT YNTRSSTKTP VASLSAAQPT ASSYNTNTAT ATPSTSQWGS STSVPQTRSS ARSTYDTNPT TSASNQYNMT PSNPPQSSSL QGFNVRPQSM AQTRSSTAAY AQQQQQHHQQ QSQAQQQQRQ QVQQQPSYNS YTSQSHTTNT QQQQHQQQQQ TQNWGYGSFG STTNASSTGY GSTGASSSAN NSYSTGTSAS SHAHNAAPAA TNAAAYSHQQ PHHRSMNLSS NTYSSHIGNE DQVLYDLLRH PGA

    DMM-2, 121,736.9 Da (1133 aa) 18 unique peptides, 250/1133 amino acids (22% coverage)

    Honda_Supplementary Fig. 7

  • D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D S ∆d

    mm -2

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D S ∆d

    mm -2

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    D S ∆d

    mm -2

    1

    2 1.6

    3 5

    0.5

    8:A6 Ψ63 8:F10

    kb 10

    probe:

    Honda_Supplementary Fig. 8

  • 8:G3probe:

    1

    2 1.6

    3 5

    0.5

    kb 10

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -1

    DS +d

    mm -1-

    HA A B

    8:G3probe:

    1

    2 1.6

    3 5

    0.5

    kb 10

    D D DS S S W

    T ∆d

    im -2

    ∆d mm

    -2

    DS +F

    LA G-

    dm m-

    2

    Honda_Supplementary Fig. 9

  • 8:G3probe:

    1

    2

    1.6

    3 5

    0.5

    kb 10

    D DS S W

    T ∆d

    im -2

    ∆d mm

    -1

    D DS S ∆d

    mm -1

    & ∆d

    im -2

    EtBr

    D DS S W

    T ∆d

    im -2

    ∆d mm

    -1

    D DS S ∆d

    mm -1

    & ∆d

    im -2

    Honda_Supplementary Fig. 10

  • 0 0 20 40

    20

    10

    15

    5

    Time (h)

    D is

    ta nc

    e (c

    m )

    WT

    ∆dmm-1 dim-5

    ∆dmm-1& dim-5

    0 0 20 40

    20

    10

    15

    5

    Time (h)

    D is

    ta nc

    e (c

    m )

    WT

    ∆dmm-1 hpo

    ∆dmm-1& hpo

    A B

    Honda_Supplementary Fig. 11

  • 0 0 20 40

    20

    10