[Fragile X mental retardation protein interacts with human NDK/Nm23-H2].
OBJECTIVE
To investigate the physiological role of fragile X mental retardation protein (FMRP) and screen the proteins interacting with FMRP in human fetal hippocampus cDNA library.


METHODS
Human fetal hippocampus cDNA library was constructed in yeast two-hybrid DAD vector pGAD10. Quality of the library was measured by picking up random colonies as templates for PCR testing. Proteins interacting with FMRP were screened by yeast two-hybrid system. Furthermore, the interaction site of FMRP was mapped in yeast.


RESULTS
The average length of inserts of the two-hybrid library was 1.5 kb, and the ratio of recombinant colonies was about 90%. Human NDK/Nm23-H2 was found interacting with FMRP. NDK/Nm23-H2 interacted with FMRP exon 1-12, as well as FMRP isoforms without exon 12, and exons 14-17. NDK/Nm23-H2 couldn't interact with FMRP exon 1-6 and exon 2-7 fragments.


CONCLUSIONS
Human NDK/Nm23-H2 can bind FMRP directly. The interaction site of FMRP is located at its exon 1-11. This interaction in vitro might alter the intracellular distribution of NDK/Nm23-H2, and even regulates the transcription and expression of FMRP.
Lou
R
R
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Mei
P C
PC
Gong
J Y
JY
Zhu
N
N
Kou
Z H
ZH
Wu
G Y
GY
Shen
Y
Y
D051860:Fragile X Messenger Ribonucleoprotein 1; D005600:Fragile X Syndrome / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D054778:NM23 Nucleoside Diphosphate Kinases; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009701:Nucleoside-Diphosphate Kinase / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D051860:Fragile X Messenger Ribonucleoprotein 1; D054778:NM23 Nucleoside Diphosphate Kinases; D009419:Nerve Tissue Proteins; D009701:Nucleoside-Diphosphate Kinase; D011506:Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C497683:FMR1 protein, human; C517938:NME1 protein, human