They are required for the male cell fates not only in soma and germline of XO males but also during the spermatogenesis period in XX hermaphrodites (6).
Homozygous mutant alleles of these genes could cause feminization of the animal (7).
CC1/2 was used to determine the high resolution limit of the diffraction data (11).
Distinct from classical PP2Cs, FEM-2 harbors an additional large N-terminal domain with no sequence similarity to any other domains known to date (9). Here, we report the crystal structure of FEM-2 and reveal that although the C-terminal domain of FEM-2 has a similar structure as several other PP2C-containing proteins, its N-terminal domain displays a new structural fold.
Results: The N-terminal domain (NTD) of FEM-2 binds to FEM-1 and FEM-3, but does not directly regulate the phosphatase activity of FEM-2.
Conclusion: The FEM-2 NTD functions as a scaffold in sex determination.
The target protein was expressed in for 45 min at 4 °C to remove cell debris.
The supernatant was applied onto a self-packaged GST affinity column (2 ml glutathione-Sepharose 4B; GE Healthcare), and contaminant proteins were removed with wash buffer (lysis buffer plus 200 m Na Cl).