Crystal structure of eukaryotic translation initiation factor eIF5B (517-970) from Chaetomium thermophilum in complex with GDP

eIF5B employs a novel domain release mechanism to catalyze ribosomal subunit joining.

PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)

Eukaryotic translation initiation factor 5B-like protein

SHMNKDNLRSPICCILGHVDTGKTKLLDKIRQTNVQEGEAGGITQQIGATYFPVEAIKQKTAVVNKDGKFEFKVPGLLII
DTPGHESFSNLRSRGSSLCNIAILVVDIMHGLEPQTIESLRLLRERKTPFVVALNKIDRLYGWKKIENNGFRESFALQNK
AVQNEFRNRLDQVKLQFAEQGFNSELFYENKNFARYVSLVPTSAHTGEGIPDMLKLIVQLCQERMASSLMYLSELQATVL
EVKAIEGFGVTIDVILSNGILREGDRIVLCGLEGPIKTNIRALLTPAPMRELRIKGQYIHHKEVKAAQGVKISAPGLEGA
IAGSRLLVVGPDDDEEELEEEVESDLQSLFSRVEKTGKGVSVQASTLGSLEALLDFLKDCKIPVANVGIGPVYKRDVMQC
GIMLEKAPDYAVMLCFDVKVDKEAQQYADENGIKIFTADIIYHLFDQFTKHMQEQLE

Chaetomium thermophilum var. thermophilum

15 % PEG 8000, 0.5 M Li2SO4, VAPOR DIFFUSION, SITTING DROP, temperature 293K

THE MISMATCH (RESIDUES 576-599) IS A WRONGLY ASSIGNMENT INTRON IN THE CHAETOMIUM THERMOPHILUM SEQUENCE. MULTIPLE SEQUENCE ALIGNMENTS WITH HOMOLOGUES FROM OTHER SPECIES SHOWED THAT THIS INTRON CORRESPONDS TO A PROTEIN SEQUENCE, WHICH IS PRESENT AND HIGHLY CONSERVED IN ALL OTHER SPECIES. IN ADDITION, ISOLATION OF THE GENE FROM A CHAETOMIUM THERMOPHILUM CDNA LIBRARY WITH APPROPRIATE PRIMERS AMPLIFIED THE GENE INCLUDING THE WRONGLY ASSIGNED INTRON.

Translation initiation, GTPase, eIF5B/IF2, Subunit joining, Ribosome, TRANSLATION

NKDNLRSPICCILGHVDTGKTKLLDKIRQTNVQEGEAGGITQQIGATYFPVEAIKQKTASFSNLRSRGSSLCNIAILVVD
IMHGLEPQTIESLRLLRERKTPFVVALNKIDRLYGWKKIENNGFRESFALQNKAVQNEFRNRLDQVKLQFAEQGFNSELF
YENKNFARYVSLVPTSAHTGEGIPDMLKLIVQLCQERMASSLMYLSELQATVLEVKAIEGFGVTIDVILSNGILREGDRI
VLCGLEGPIKTNIRALLTPAPMRELRIKGQYIHHKEVKAAQGVKISAPGLEGAIAGSRLLVVGPDDDEEELEEEVESDLQ
SLFSRVEKTGKGVSVQASTLGSLEALLDFLKDCKIPVANVGIGPVYKRDVMQCGIMLEKAPDYAVMLCFDVKVDKEAQQY
ADENGIKIFTADIIYHLFDQFTKHMQEQLE