(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.
Fig. 2. CAR-1 and CGH-1 associate together in a RNA-dependent complex. (A)
Immunoprecipitation of endogenous CGH-1-associated proteins from one-day-old
adult hermaphrodites, identified by mass spectroscopy. (B) Each CAR-1 ortholog
includes a Sm-like domain within a characteristic conserved region (black) and
a variable number of Arginine-Glycine-Glycine (RGG) triplets or functional
equivalents (vertical bars) (Birney et al.,
1993). An additional conserved region (shaded) contains a FDF
domain, a sequence motif of unknown function
(Anantharaman and Aravind,
2004). The percentage identity (Id) and similarity (Sim) to C.
elegans CAR-1 within these regions was determined by pair-wise BLAST.
CAR-1, C. elegans (NP_493254); Hs, H. sapiens (NP_056393);
Tral, D. melanogaster (AAF49905); Scd6p, S. cerevisiae
(NP_015454). (C) Interaction between CAR-1 and CGH-1 requires RNA.
Co-immunoprecipitation of CAR-1 and CGH-1 from C. elegans extracts
(lanes 2 and 6) was abolished by RNase A treatment (lanes 3 and 7). Lanes 4
and 8 show control incubations. (D) CAR-1 and CGH-1 are detected specifically
in the germline. Lysates from one-day-old adult wild-type worms (N2) and
germline-deficient glp-4(bn2) hermaphrodites
(Beanan and Strome, 1992) were
analyzed by western blotting with CAR-1 and CGH-1 antisera, using Tubulin as a
control. The CAR-1 antibody recognized a single germline-specific species of
45 kDa, larger than the predicted size of 37.6 kDa. (E) Diminished CAR-1
protein levels in car-1(RNAi) worms. Protein extracts from wild-type
(N2) or car-1(RNAi) one-day-old adults were analyzed by western
blotting.
