[Screening of proteins interact with FMR1 by yeast two-hybrid system].
OBJECTIVE
To establish yeast two-hybrid system for screening of protein(s) interacted with the fragile X metal retardation protein (FMRP).


METHODS
Fragment of exon 11 to 15 of the FMR1 cDNA was recombined with DNA-binding domain of the pBTM116 vector as bait to screen a mouse embryo cDNA library.


RESULTS
Thirteen clones were confirmed to be able to specifically interact with FMRP bait. Sequence analysis showed that 12 clones are overlapping ones containing cDNA fragments of the mouse ubiquitin-conjugating enzyme gene (mUBC9).


CONCLUSIONS
The interaction between UBC9 and FMRP is supported both by similarity search of the amino acid sequences of the mouse and human UBC9 and by expression characteristics of the two proteins. The biological significance of the interaction is to be further studied.
Chen
Y
Y
Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China.
Sittler
A
A
Yu
M
M
Bardoni
B
B
Wu
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics; D004622:Embryo, Mammalian; D005091:Exons; D051860:Fragile X Messenger Ribonucleoprotein 1; D015723:Gene Library; D008025:Ligases / Q000737:chemistry / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics*; D016601:RNA-Binding Proteins; D020539:Sequence Analysis, Protein; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9
D004268:DNA-Binding Proteins; D051860:Fragile X Messenger Ribonucleoprotein 1; D008025:Ligases; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9; C497684:Fmr1 protein, mouse