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Supplementary Information A. Homology between Daysleeper and other hAT transposases
Daysleeper
Tam3
Ac
Tag1
1
1
1
1
----------------------------------------------------MEVYNDDT
------------------MDTSNIQPATTEDYIHNFNMGVGFSVWDTTLGIGPHVGNYQQ
MTPPVGNNPPSGSAIRLAKLMSTTRAPSTRKTNSVFSA-YAQGLKRKAEASSSRIQNVRA
--------------------METEHDENYEDIAAANRS----IRGKSDIAWSYVIQSKDE
Daysleeper
Tam3
Ac
Tag1
9
43
60
37
EMR-SPETQPIK--ETALEVYNDTAEIRSPETQPIEETALEVYNDT-EMVSPETQPIKRR
EHVFGPGVGPSNNNPAVELGPHVGTSHHDTSANMAGETATGGASSA-VGSDDDAAEGDNA
RAR-GHGCGRTS--PSSSTAEAERHFIQSVSSSNANGTATDPSQDD-MAIVHEPQPQPQP
KGKTVLECAFCH--KKKRGGGINRMKHHLAGVKGNTDACLKISADIRFKIVNALKEAENK
Daysleeper
Tam3
Ac
Tag1
65
102
116
95
* *
* *
* *
*
KKKSMVWEHFTIEAVEPNCRRAFCKGCN-QSFAYSNGN-KVAG--TSHLKRHIFKGTCPA
EMDEDEIPDTPVEQMASTSRPPKKTKKA----TVWKWFSKVTG----SNWAQCLL--CPT
QPEPQPQPQPEPEEEAPQKRAKKCTSDVWQHFTKKEIEVEVDGKKYVQVWGHCNFPNCKA
KKQNIVLDDTNLDGPEIEDVDGDDIRVD----VRPSQKRKKQG-----IDLHDYF----K
-----------------------------------------------------*
Daysleeper
Tam3
Ac
Tag1
121
152
176
142
Daysleeper
Tam3
Ac
Tag1
172
207
235
181
Daysleeper
Tam3
Ac
Tag1
231
267
294
238
LI-----HTHDNDNNPLMSTPYTPK-TDTPRRRYRSQ---NNASPYVAFNQDKCRQEIAK
RYSHKTGCGTGTLTRHLTAKHKNRD-MDAPDMQRQPD---GTMAP-WRYDQNYMRICLAQ
KYRAEGHHGTSGFRNHLRTSHSLVK-GQLCLKSEKDHGKDINLIEPYKYDEVVSLKKLHL
R---------G---VHDQTQPSIKACMQSKERIHAVD----MSVALWFYD-----ACIPM
*
**
* *
—
MIIMHDYPLHMVQHPGFVSFVQ-SIQPHFDAVSFNNVQGDCVATYLAEKQNVMKSLEGIP
FIVQNELPFSFAQNELFENFLQKAVQCKFKKISRATCFRDGVKQYEKEIIVLRNEFKNFN
AIIMHEYPFNIVEHEYFVEFVK-SLRPHFPIKSRVTARKYIMDLYLEEKEKLYGKLKDVQ
NAVN--SPLFQPMMSMVASMGHGYVGPSYHAL-RVGLLRDAKLQVSLIIDKFKSSWASTG
*
A
—————————————
GRFCLTLDFWTSKLTLGYVFITAHYIDSDWKIQKKLLNVLMESYPEADEALSLAVANCVS
GRISLTSDLWQGSGSYHFSCITAHWIDKDWIMRKRIIEFAQLDSPHNGDCIRDATMSSLN
SRFSTTMDMWTSCQNKSYMCVTIHWIDDDWCLQKRIVGFFHVEGRHTGQRLSQTFTAIMV
--CTLMADGWKDTRQRPLINFLV-YCPKGITFLKSVDASDIYASAENLCNLFAELVGIIG
291
327
354
295
B
—————————————
EWGLEGKLFNVTFNHPASNS-AVENIRPQLCIKNP-GILDGQLVIGNCVARTFGSLAKDV
YWGIKDKIMSISLDNASNNVNAIKSLKPAMN-----LILGGQLFHVRCICHILHLCVKDG
KWNIEKKLFALSLDNASANEVAVHDIIEDLQDTDSNLVCDGAFFHVRCACHILNLVAKDG
SENIVHFVTDSAPNYKAAGKLLVEKFPTIAWSPCSAHCINLILEDVAKLPHVHHIVRR--
Daysleeper
Tam3
Ac
Tag1
349
382
414
353
C
——————————
LEKGKDVIKNIRDSVKHVKTSESHEERFTELKEQLQVPSEKVLSLDDQTQWNTTYMMLVA
LSVLIQSIDRIRVCLSHINRYPPRVQAFNTVCETHGMP-IKHIYLDVPHRWNATYRMLIE
LAVIAGTIEKIKAIVLAVKSSPLQWEELMKCASECDLDKSKGISYDVSTRWNSTYLMLRD
MSKVTIFVYNHKPALNWVRKR-SGWREIIRPGETRFATTFIALQSLYQHKEDLQALVTSA
Daysleeper
Tam3
Ac
Tag1
409
441
474
412
ASELKE--VFSCLDTADP--DYKKPPSAEDWRHVEALCTFLKPLFEAVSTLQSTGNPSAV
AKPYSEPITFFCHRSLGP---NSIL--ADDWNICDILVPYLVYFEEFTKIMSSCYTPTSN
ALYYKP--ALIRLKTSDPRRYDAICPKAEEWKMALTLFKCLKKFFDLTELLSGTQYSTAN
DPELKQ--LFKTSKAKVA---KSVILDERMWNDCLIIVKVMTPIIRLLRICDADEKPSLP
Daysleeper
Tam3
Ac
Tag1
465
496
532
467
TFFHEVWKTQSDLSRAIAGEDPFVTGIAKTMQEKVDKYWRDCSLVLAMAVVMDPRFKMKL
IMLLYMVSVVRLFHQHRNHAT--LKNIIKEMEKKWVKYYKKVPNVCILSSCLDPRVRLIG
LFYKGFCEIKDLIDQWCVHEKFVIRRMAVAMSEKFEKYWKVSNIALAVACFLDPRYKKIL
YVYEGMYRARLGIKNIFQEKETLYKPYTNIIDRRWDRMLR--HDLHAAAYYLNPAFMYDQ
Daysleeper
Tam3
Ac
Tag1
Daysleeper
Tam3
Ac
Tag1
525
554
592
525
VEFSFS-KIFGEDAGKN----IKTVDDGIHELFTEYMALP--SPQNTTSEGGKADG---TLELLE-KYHSALNNVYNG--NEERNRILQLLYSLYDMYAPSTDMDESPTNASRGSG--IEFYMK-KFHGDSYKVH----VDDFVRVIRKLYQFYSSCSPSAPKTKTTTNDSMDDTLME
PTFCEKPEVMSGLMNLFEKQKNDSKTKLFQELR-VYREREGSFSLDMALTCSKTSQP---
574
608
647
581
D
———————————
——————
--LSDFDTYIMETTG--QNLK--SELDQYLDETLLPRVQ-EFDVLDWWKQNKLKYPTLSK
--FSIFDELLSNQQSNQPSVGNYTEIHLFVQKPPQKFDK-DFDILKWWRQNESLTPVLAR
NEDDEFQNYLHELKDYDQVES--NELDKYMSEPLLKHSG-QFDILSWWRGRVAEYPILTQ
--DEWWRYFGHDAPNLQKMAI--RILSQTASSSGCERNWCVFERIHTKKRNRLEHQRLND
Daysleeper
Tam3
Ac
Tag1
627
665
704
637
E
F
———————————————————— ————————————————————————————
MARDILSIPVSAAAF-DYVFD-MEPREMDEYKTSLRPETVEALICAREWLLESN-----IARDLLSSQMSTVAS-ERAFS-AGHRVLTDARNRLKPGSVKFCMIWKDMLDQQYR----IARDVLAIQVSTVAS-ESAFS-AGGRVVDPYRNRLGSEIVEALICTKDWVAASRKGATYF
LVFVHYNLRLQHRSKRKRSYDPVDYESIDKTEFWVVEE-EEAGELEYDELENALAEEYP-
Daysleeper
Tam3
Ac
Tag1
679
718
762
695
----------ASSSAAAQNASATIKSEA------------------------EKTLRKVETVLAMDGEEDNEDATMPYDFTNIH------PTMIGDLEVLDSVIAAATNHENHMDEDEDAIEFSKNNEDVASGSSP
----KDLEDTNPETSNDFDEDFTLPPEEDVWNDEEDNRD-------
Daysleeper
Tam3
Ac
Tag1
AAT77879
XP_475955
XP_475956
At1g15300.1
Tam3.1
XP_473603
XP_473986
XP_507160
XP_463295
Daysleeper N
F96831
Alignment of Daysleeper with the transposases from the mobile elements Activator (Ac) from maize, Tam3
from Antirhinum majus and Tag1 from Arabidopsis thaliana. The BED-type zinc finger domain in
Daysleeper is indicated with a dashed line with the mutations that abolished Daysleeper binding to Kubox1
multimers indicated with an asterisk (*). Amino acid residues shown to be essential for Ac transposition are
indicated below the Ac sequence by a bold asterisk (*). The hAT conserved domains 1 (A-F) are indicated.
The conserved Ac dimerization domain is located on the Ac sequence between the residues 646-754
(Essers, L., Adolphs, R.H. & Kunze, R. (2000) A highly conserved domain of the maize Activator
transposase is involved in dimerization. Plant Cell 12, 211-233). The amino acids important for
dimerization (I704, V708, L709, R733 & L744) are also well conserved. The alignment was performed with
the Clustal W program.
A BLAST search using the N terminal region of Daysleeper identified several proteins from other plant
species sharing high homology. A phylogentic tree of the N terminal regions of these proteins is shown.
The proteins AAT77879, XP_475955, XP_475956, XP_473603, XP_473986, XP_507160, and
XP_463295 originate from rice, Tam3.1 originates from Antirhinum majus and At1g15300.1 and F96831
come from Arabidopsis. The N terminus of transposases tends to be less conserved, as is clear from the
alignment above. Thus, proteins sharing homology in this region are more likely to have a conserved
function in the plant.