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Zeste maintains repression of Ubx transgenes: support for a new model of Polycomb repression

Man-Wook Hur1, Jeffrey D. Laney2, Sang-Hack Jeon3, Janann Ali4 and Mark D. Biggin5,*

1 Department of Biochemistry and Molecular Biology, Yonsei University School of Medicine, 134 ShinChon-Dong, SeoDaeMoon-Ku, Seoul, Korea 120-752
2 Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA
3 Department of Biology, Kon-Kuk University, Seoul, Korea 143-701
4 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA
5 Genome Sciences Department, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Mail stop 84-171, Berkeley, CA 94720, USA



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  		Fig. 1. Transgenic analysis of Ubx proximal promoter mutations. Cartoons of promoter constructs tested are on the left and indicate the mRNA start site at +1 (arrow), the number of factor binding sites present at the proximal promoter (P) of each construct, and the position of the ß-galactosidase reporter gene (lacZ). Representative pictures of stage 13-14 embryos showing the pattern of ß-galactosidase expression from each transgene are to the right. The normal anterior and posterior boundaries of Ubx expression are marked with arrowheads. (A) 22UZ Native; (B) 22UZ Deletion; (C) 22UZ GAGA; (D) 22UZ ZESTE; (E) 22UZ NTF-1; (F) 22UZ GAGA/NTF-1; (G) 22UZ ZESTE/NTF-1.

 


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  		Fig. 2. Comparison of transgene expression in wild type and zeste mutant embryos. (A,B) 22UZ GAGA. (C,D) 22UZ ZESTE. (E,F) 22UZ NTF-1. (G-K) 22UZ ZESTE/NTF-1. Expression in wild-type embryos is shown on the left (Wt). Expression in zeste mutant embryos is shown on the right (Zmt). Essentially identical results were observed in zae(bx) and zv77h embryos for all constructs (H,I and M.-W. Hur, unpublished). All embryos are stage 13-14, except J and K, which are stage 11.

 


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  		Fig. 3. Derepression of 22UZ Zeste and 22UZ GAGA transgenes in Pc3 mutant embryos. (A,B) 22UZ GAGA. (C,D) 22 UZ Zeste. Expression in wild-type embryos is shown on the left (Wt). Expression in Pc3 embryos in shown on the right (Pc3). The arrowheads mark the anterior and posterior boundaries of Ubx expression in wild-type embryos. The increased expression to the left of the anterior arrowhead in Pc3 homozygous embryos shows the derepression of the transgenes. All embryos were stained for the same length of time.

 




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