Proteins co-expressed in bacteria, purified by virtue of His tag, and the heterodimer purified by gel filtration.

Proteins expressed and purified from bacteria.

Smn and Gem2 form a heterodimer in vitro.

Components mixed and the complex purified by gel filtration.

Smn interacts with Gem2, SmD1, SmD2, SmE, SmF and SmG to form a 7S Sm core complex intermediate.

Gel filtration of the complex indicates a molecular mass consistent with a monomeric stoichiometry of each constituent.

Proteins expressed and purified from bacteria.

Purified recombinant D. melanogaster proteins Gem2, Smn and icln were mixed with human SmD1, SmD2, SmE, SmF and SmG to reconstitute the 8S spliceosome assembly intermediate.

Electron microscopy 3D reconstruction was used to position those elements of the 8S complex that could not be traced in the crystallographic electron density.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein.(Pairwise.)

MORATORIUM:This data was made available \'pre-publication\' and, as such, its use is subject to the specific limitations described in FBrf0235788 for the period of 12 months. Moratorium expires on June 13, 2018.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein.(Pooled)

MORATORIUM:This data was made available \'pre-publication\' and, as such, its use is subject to the specific limitations described in FBrf0235788 for the period of 12 months. Moratorium expires on June 13, 2018.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein.(Pairwise.)

MORATORIUM:This data was made available \'pre-publication\' and, as such, its use is subject to the specific limitations described in FBrf0235788 for the period of 12 months. Moratorium expires on June 13, 2018.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein.(Pooled)

MORATORIUM:This data was made available \'pre-publication\' and, as such, its use is subject to the specific limitations described in FBrf0235788 for the period of 12 months. Moratorium expires on June 13, 2018.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Smn and Gem2 behave as a distribution of tetramers and octamers of Smn-Gem2 heterodimers in solution. Mutations Y204H, Y204R, Y208A shift the Smn-Gem2 complex towards smaller species while G206S results in monomeric Smn-Gem2 heterodimers.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

Smn and Gem2 form oligomers of Smn-Gem2 heterodimers. Sedimentation coefficients are temperature sensitive with shift to smaller species at lower temperatures. Mutations Y208H and Y204R also shift the Smn-Gem2 complex towards smaller species.

Interaction in vitro; proteins produced as recombinant fusion proteins in a bacterial system.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

Smn and Gem2 behave as a distribution of tetramers and octamers of Smn-Gem2 heterodimers in solution.