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Michael Leunig A putative transcription co-repressor complex

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Presentation on theme: "Michael Leunig A putative transcription co-repressor complex"— Presentation transcript:

1 Michael Leunig A putative transcription co-repressor complex
regulates floral organ identity and shape by Zhongchi Liu, University of Maryland, USA Dr. Seuss Gary Larson Michael Leunig

2 Floral homeotic gene AGAMOUS (AG) specifies stamen and carpel identity
WT whorl 1 whorl 2 whorl 3 whorl 4 ag-1 AG expression in WT ag-1 Yanofsky et al., Nature, 1990; Drews et al., Cell, 1991

3 leunig (lug) mutation results in ectopic expression of AG
wild-type lug lug-1 H S X B E3 GUS 2.98kb E2 Liu and Meyerowitz, Development (1995); Busch et al., Science (1999); Deyholos and Sieburth, Plant Cell (2000)

4 Structural Similarity to Transcription Co-repressors
1 931 LUG (Arabidopsis) Q-rich (89-184, ) LUFS 7 WD 1 787 LUH (Arabidopsis) Q-rich LUFS 7 WD 1 706 Tup1 (Yeast) Ssn6 Binding Q-rich (97-118, ) 7 WD 1 716 Groucho (Drosophila) Q-rich Ser-pro rich 7 WD Conner and Liu, PNAS, (2000)

5 The repression of a-cell-specific genes in S. cerevisiae a cells
Tup1 Ssn6 a-cell specific genes a 2 MCM1 The repression of AG in first two whorls of Arabidopsis flower LUG ? ? AG exon ? AG second intron

6 seuss (seu) resembles lug and enhances lug
lug seu

7 AG mRNA expression WT lug seu

8 SEU encodes a Q-rich protein with a conserved dimerization domain
Q-rich domain LIM Interacting domain Frameshift (seu-2) Q to stop (seu-1) 1 877 SEUSS NLS 1 933 Rice EST 81% 1 748 Arabidopsis Homolog 55% 1 384 Zebra fish LIM domain- binding factor (Ldb1) 22% LID 1 375 Mouse Ldb1 21% LID

9 LUG LUFS domain interacts with full length SEU
LUG-DB Constructs LUFS LUFS Q-rich (89-184, ) LUFS Q-rich (89-184, ) 7 WD Q-rich (89-184, ) 7 WD

10 1. 2. 3. 4. 5. 6. Reporters 5x Gal4UAS CaMV 35S GUS 3’NOS LUC GAL4DB
LUG SEU Effectors 3. 4. 5. 6. 2 4 6 8 10 12 14 16 18 GUS/LUC LUG-BD (1+2+4) SEU-BD (1+2+5) SEU No Effector 1+ 2 GAL4BD 1+2+3 1+2+ effector

11 LUG SEU ? AG exon ? AG second intron

12 larson-1 (lsn-1) enhances lug
at 20oC ap2-2 lsn-1 lug-8 lsn-1 lug-8 lsn-1 lug-8

13 lsn-1 is temperature-sensitive
lsn-1 at 20 oC lsn-1 at 29 oC

14 Ectopic AG expression in lsn-1 at 29oC
AG second intron H S X B E3 GUS 2.98kb E2

15 LSN encodes a BEL1-like homeodomain protein
575 349 414 100% lsn-1 P>L lsn-2 R>L 391 456 85% BEL1 610 193 SKY domain BEL domain Homeodomain

16 LSN Belongs to The TALE Superclass Homeodomain Protein
Helix 1 Helix 2 Helix 3 Cons. RELSKKRKRGKLPKEATQILRAWWPQHLKNPYPSEEEKEALAEETGLTLKQINNWFINARRRHWKPMIDMQ IRO VTYDLAARRKNATRESTSTLKAWLNEHKKNPYPTKGEKIMLAIITKMTLTQVSTWFANARRRLKKENKMTW PBC REQSRRRKRRNFNKQATEILNEYFYSHLSNPYPSEEAKEELAKKCGITVSQVSNWFGNKRIRYKKNIGKFQ KNOX LEFSKKKKKGKLPKEARQQLLEWWKGHYKWPYPTESDKISLAESTGLDPKQINNWFINQRKRHWKPSEHMQ CUP RRKQN-SGRSNLPKETVSILNTWLLNHLNNPYPTQQEKRELLIKTGLTKIQLSNWFINVRRRKIFSDYYTL TGIF GSGKR-RRRGNLPKESVQILRDWLYEHRYNAYPSEQEKALLSQQTHLSTLQVCNWFINARRRLLPDMLRKD MEIS KDKKRQKKRGIFPKVATNIMRAWLFQHLTHPYPSEEQKKQLAQDTGLTILQVNNWFINARRRIVQPMIDQS M-ATYP VVSPAVGCRNLSEDLPAYHMRKHFLHTLDNPYPTQEEKEGLVRLTNESTARVGSWFINARRRSGWSHILKK BEL HPQHAWRPQRGLPERSVSVLRAWLFEHFLHPYPKDSDKHMLARQTGLSRNQVSNWFINARVRLWKPMVEEM Classes L L LSN HHAPVWRPHRGLPERAVTVLRAWLFDHFLHPYPTDTDKLMLAKQTGLSRNQVSNWFINARVRVWKPMVEEI BELL1 ---HPWRPQRGLPERAVTTLRAWLFEHFLHPYPSDVDKHILARQTGLSRSQVSNWFINARVRLWKPMIEEM ATH1 KNHQIWRPQRGLPEKSVSVLRNWMFQNFLHPYPKDSEKHLLAIRSGLTRSQVSNWFINARVRLWKPMIEEM AAD EAWRPQRGLPERSVNILRAWLFEHFLHPYPSDADKHLLARQTGLSRNQVSNWFINARVRLWKPMVEEM AAF18625 HHSNAWRPQRGLPEKAVSVLRSWLFEHFLHPYPRDLDKVMLAKQTGLTKSQVSNWFINARVRMWKPLVEEL AAK EAWRPQRGLPERSVNILRAWLFEHFLNPYPSDADKHLLARQTGLSRNQVSNWFINARVRLWKPMVEEM AAK43836 PSNNAWRPQRGLPERAVSVLRAWLFEHFLHPYPKDSDKHMLAKQTGLTRSQVSNWFINARVRLWKPMVEEM AAL HTWRPQRGLPDSSVLVLRAWLFEHFLHPYPKDSDKIMLARQTGLSRGQVSNWFINARVRLWKPMVEEM AAL AWRPQRGLPENSVSILRAWLFEHFLHPYPKESEKIMLSKQTGLSKNQVANWFINARVRLWKPMIEEM NP QQQHIWRPQRGLPERAVAVLRAWLFDHFLHPYPTDSDKQMLATQTGLSRNQVSNWFINARVRLWKPMVEEI NP QAWRPQRGLPENSVLILRAWLFEHFLHPYPKDSDKIMLARQTGLSRGQVSNWFINARVRLWKPMVEEI ***:****: :* :** *:*::****** * :* :*: ::**:*:**:*********:****:* Arabidopsis Homologues

17 Structure of Hox B1-Pbx1 heterodimer bound to DNA
Piper,D.E., Batchelor,A.H., Chang,C.P.,Cleary,M.L. and Wolberger,C. Cell 96 (4), (1999)

18 Gal2p lacZ Gal7p ADE2 Gal1p HIS3
DB AD pJ69-4A yeast host LUG LSN D81 (three reporters) Gal2p lacZ LUG + Gal7p ADE2 + LSN Gal1p HIS3 + + LUG LSN D3 575 349 414 HD 81 pVA3 pTD1 LUG SEU

19 Binding of LSN to AG second intron
2985 AG GST-LSN GST

20 In whorls 1 and 2 of a flower
A proposed model on how LUG and SEU repress AG In whorls 1 and 2 of a flower LUG SEU AP1? AG exon LSN AG second intron

21 seu and lug mutants develop narrow floral organs

22 SEU and LUG regulate organ shape independently of AG
ag-1 seu-1 ag-1 ag-1 seu-1 lug-1 seu-1 lug-1 ag-1 seu-1 lug-1

23 The narrow organ shape may result from a loss of organ polarity
ag-1 ag-1 seu-1 lug-1 PROBE PHABULOSA (Adaxial) YABBY1 (Abaxial) See poster # 14 by Robert Franks

24 In whorls 1 and 2 of a flower
Organ shape LUG In whorls 1 and 2 of a flower SEU ? X PHB/YAB Floral organ identity LUG SEU AP1? AG exon LSN AG second intron

25 ACKNOWLEDGEMENT Postdoctoral fellows Joann A. Conner Robert G. Franks
V.V. Sridhar Graduate students Alex Xiaozhong Bao Anandkumar Surendrarao Jayashree Sitaraman Black Chunxin Wang Joshua Levin, Syngenta John Bowman, UC Davis Steve Henikoff, FHCRC Our work is funded by USDA, DOE, NSF and a NIH postdoctoral fellowship to B. Franks

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