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Fig. 4. Proteolytic cleavage converts Cvl2 from an anti- to a pro-Bmp
factor. (A) Western blot with anti-Myc antibody after SDS-PAGE with
embryonic protein extracts of cvl2-myc mRNA-injected embryos. Under
reducing conditions [+mercaptoethanol (ME)], full-length Cvl2 and a C-terminal
cleavage fragment are detectable. Under non-reducing conditions (without ME),
the cleavage products remain associated, and only the high molecular weight
band is detectable. (B) Illustration of Cvl2 proteins encoded by the
different mRNAs. The protein encoded by cvl2-WT is either cleaved,
with the two fragments remaining associated via disulfide bounds of thus far
unidentified residues (top), or uncleaved, resembling Cvl2-CM (lower panel).
(C) Western Blot showing that Cvl2-CM is not cleaved in embryos, and
that Cvl2-N is readily synthesized. (D-G) Lateral views of 32 hpf
embryos; (H-O) animal views, dorsal rightwards, after in situ
hybridization at 80% epiboly for the ventral marker eve1 (H-K) or the
dorsal marker chordin (chd; L-O)
(Schulte-Merker et al., 1997).
Injection of cvl2-wt mRNA leads to both weakly ventralized (V1 in D
and right embryos in H,L) and weakly (C1 in D) or moderately dorsalized
embryos (C2 in D and left embryos in H,L). Injection of cvl2-CM mRNA
causes strong dorsalization (F,J,N), while cvl2-N mRNA causes
ventralization (G,K,O). Arrows in I,K,M,O indicate borders of the
eve1 or chd expression domains. Ratios of obtained
phenotypes were: (D-G) see Table
1; (H) dorsalization, 17/44; wild type, 22/44; mild
ventralization, 5/44; (J) 52/55; (K) 21/43; (L) dorsalization, 21/56; wild
type, 29/56; mild ventralization 6/56; (N) 45/48; (O) 21/36.
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