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First published online March 24, 2005
doi: 10.1242/10.1242/dev.01776

Development 132, 1885-1893 (2005)
Published by The Company of Biologists 2005
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Components of the transcriptional Mediator complex are required for asymmetric cell division in C. elegans

Akinori Yoda1,2, Hiroko Kouike1,3,4, Hideyuki Okano1,3,4 and Hitoshi Sawa1,5,6,7,*

1 Division of Neuroanatomy, Osaka University Graduate School of Medicine, Kobe University, Kobe 650-0017, Japan
2 Department of Genome Sciences, Graduate School of Medicine, Kobe University, Kobe 650-0017, Japan
3 CREST, Japan Science and Technology Corporation, Japan
4 Department of Physiology, Keio University School of Medicine, Tokyo 160-8582, Japan
5 PRESTO, Japan Science and Technology Corporation, Japan
6 Division of Bioinformation, Department of Biosystems Science, Graduate School of Science and Technology, Kobe University, Kobe 650-0017, Japan
7 Laboratory for Cell Fate Decision, Riken, Center for Developmental Biology, Kobe 650-0047, Japan



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  		Fig. 1. Abnormal T-cell lineages in let-19 and dpy-22 mutants at the L1 stage. The fates of cells (H, hypodermal; N, neural) were determined by nuclear morphology. The number of animals that showed a given lineage is indicated below the diagrams.

 


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  		Fig. 2. Asymmetric expression of POP-1 in the T-cell division is not affected by let-19 and dpy-22 mutations. Expression of GFP::POP-1 in L1 larvae of wild-type (A), lin-17(n3091) (B), let-19(mn19) (C) and dpy-22(os38) (D). Anterior is towards the left, ventral towards the bottom. The daughters of the T cells are indicated.

 


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  		Fig. 3. Symmetric expression of TLP-1 in let-19 and dpy-22 mutants after the T-cell division. Expression of tlp-1::GFP in L1 larvae of wild-type (A), lin-17(n3091) (B), let-19(mn19) (C), dpy-22(os38) (D) or lin-17(n3091); let-19(mn19) (E). Anterior is towards the left, ventral towards the bottom. The daughters of the T cells are indicated.

 


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  		Fig. 4. Molecular cloning of let-19 and dpy-22. Genetic maps of the let-19 (A) and dpy-22 (B) loci with rescuing cosmids. Structures of the genes and rescuing constructs are shown with the coding regions in gray and the Q-rich domain in dpy-22 hatched. The molecular lesions of the mutations are indicated. The sop-1-class mutations of dpy-22 are from Zhang and Emmons (Zhang and Emmons, 2000Go). The sy622 and sy655 mutations are from Moghal and Sternberg (Moghal and Sternberg, 2003Go). The total lengths of the protein products are indicated on the left. (C) Protein sequence comparisons of the C-terminal regions of MED13 homologs from C. elegans (Ce), human (Hs), mouse (Mm), rat (Rn), D. melanogaster (Dm), D. discoideum (Dd), S. pombe (Sp) and S. cerevisiae (Sc). The consensus sequence (Cons) is indicated in the top row. The numbers indicate positions in the complete peptide sequences. Black and gray backgrounds indicate identical or similar amino acids, respectively, in at least four aligned sequences. Amino acids considered similar are R/K/H, S/T, I/L/V/M, E/D, Q/N and F/Y/W. Stop signals are indicated by asterisks. The mutation site (R2834stop) of let-19(os36) is indicated in italics.

 


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  		Fig. 5. Symmetric expression of LET-19 and DPY-22 in the T-cell division. Expression of dpy-22::GFP (A,B) and let-19::GFP (C,D) in the T cell. The T cell is in telophase in B and D. Anterior is towards the left, ventral towards the bottom.

 


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  		Fig. 6. Association of LET-19 with SUR-2 and LET-425 in vivo. Nuclear extract (NE) was prepared from sur-2 mutant animals expressing only HA-tagged SUR-2 or both GFP-tagged LET-19 and HA-tagged SUR-2. Nuclear extracts and immunoprecipitation (IP) with anti-Flag (F) and anti-GFP (G) antibodies were analyzed by immunoblotting using antibodies against GFP, HA and LET-425.

 




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