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Fig. 2 The Defective Gene in sunrise Is cdc73
(A) The sunrise locus was mapped on chromosome 2 between the microsatellite marker z19837 and z13475. The cdc73 gene is located within this region.
(B) The morphology of wild-type, sun mutant, and rtf1-/- mutant at 36 hpf. Injection of cdc73 mRNA completely (12 out of 12 embryos) rescued the morphology of sun mutants.
(C) DNA sequence chromatograms showing the C → T transition at +1507 in cdc73 coding region in sun homozygous mutant, leading to a premature stop codon (in the red box).
(D) Alignment of the C-terminal CDC73 protein sequence from Drosophila, human, and zebrafish. The nonsense mutation in sun mutant is indicated by a red arrow.
(E) Western blot showing the loss of CDC73 protein in sun mutant. Protein was extracted from 36 hpf embryos. Red asterisk: zebrafish CDC73 (∼64 kDa). Black asterisk: nonspecific bands. Actin is used as a loading control.
(F) Morpholino (MO)-mediated knockdown of cdc73 completely rescues globin expression in mon (22 out of 22 embryos), whereas 5 bp mismatch control morpholino has no rescue (0 out of 13 embryos).
Reprinted from Cell, 142(1), Bai, X., Kim, J., Yang, Z., Jurynec, M.J., Akie, T.E., Lee, J., LeBlanc, J., Sessa, A., Jiang, H., DiBiase, A., Zhou, Y., Grunwald, D.J., Lin, S., Cantor, A.B., Orkin, S.H., and Zon, L.I., TIF1gamma controls erythroid cell fate by regulating transcription elongation, 133-143, Copyright (2010) with permission from Elsevier. Full text @ Cell