Fig. 7. CSN5 mutations activate a mei-41-dependent meiotic
checkpoint. (A) In wild type, Grk protein is detectable in region 2a of the
germarium and is restricted to the oocyte from region 3 onwards. (B) In
CSN5ex21/CSN5L4032 germaria, Grk is
undetectable. (C) Loss of mei-41 restores Grk expression in
CSN5ex21/CSN5L4032 heterozygotes. (D,E)
CSN5 mutations affect the mobility of Vasa protein. (D) An anti-Vasa
western blot of ovarian extracts shows retarded Vasa migration in
CSN5L4032 germline clones. (E) An anti-Vasa western blot
of ovarian extracts from several CSN5 allelic combinations. In wild
type, Vasa migrates as a single 72 kDa band. Vasa migrates as two bands in
CSN5ex13/CSN5L4032 and
CSN5ex21/CSN5L4032. Notice, that in the
stronger combination (CSN5ex21/CSN5L4032) more
Vasa protein migrates slowly than in the weaker combination
(CSN5ex13/CSN5L4032). Furthermore, in
CSN5ex13/CSN5L4032, Vasa is fully restored to
normal mobility by removal of one dose of mei-41, while
CSN5ex21/CSN5L4032 requires homozygous
mei-41 for normal Vasa mobility. Removing one dose of
mei-W68 is sufficient to restore Vasa mobility in both hypomorphic,
CSN5-mutant combinations. A second mei-W68 allele
(mei-W681) had a weaker effect. As a heterozygote with the
CSN5-mutant combinations, it gave only a partial rescue of Vasa
mobility, although the mei-W681/mei-W68k05603
combination resulted in full rescue of Vasa mobility (data not shown).