Neural Development - BioMed Central

More documents

Recommendations

Info

Neural Development 2009, 4:1 http://www.neuraldevelopment.com/content/4/1/1 A NM23-H8 NM23-X6 NM23-H6 NM23-X1 NM23-X2b NM23-X2a NM23-H2 NM23-H1 NM23-H3 NM23-X3 NM23-H4 NM23-X4 NM23-H5 NM23-X5 NM23-H7 NM23-X7 NM23-X8 B NM23X4 NM23H4 NM23X1 NM23X2 NM23X3 NM23X4 NM23H4 NM23X1 NM23X2 NM23X3 NM23X4 NM23H4 NM23X1 NM23X2 NM23X3 NM23X4 NM23H4 NM23X1 NM23X2 NM23X3 MNFL-RRGII-AFRSAASAVPGRSECVLYSSVAGGCE----RTLIAVKPDGVQRRLVGEI 54 MGGLFWRSALRGLRCGPRAP-GPSL--LVRHGSGGPSWTRERTLVAVKPDGVQRRLVGDV 57 -----------------------------------MAANKERTFIAIKPDGVQRGLIGDI 25 -----------------------------------MAANKERTFIAIKPDGVQRGLMGDI 25 -------------------MICLVLTIFAHIFPSAWTGINERTFLAIKPDGYQRRLLGEI 41 ** * **** * * * IKRFEQRGFTLVGLKLLQASEGILAEHYHDLRRKPFYPALLRYMASGPVVAMVWEGHNVV 114 IQRFERRGFTLVGMKMLQAPESVLAEHYQDLRRKPFYPALIRYMSSGPVVAMVWEGYNVV 117 IKRFEQKGFRLVAMKFQQASQDLLRQHYIDLKDRPFYPGLVEYMSSGPVLAMVWEGLNVV 85 IKRFEQKGFRLVAMKFLQASQDLLRQHYIDLKDRPFYPGLVEYMNSGPVLAMVWEGLNVV 85 IRRFEKKGFRLVAMKIVQASEKLLKQHYIALQDKPFYDRLVKYMGSGPVVAMVWQGLDVV 101 * *** ** ** * ** * ** * *** * ** **** **** * ** RTSRGMVGDTDSSQAKPGTIRGDFSVHISRNVIHASDSVEGAEREISLWFHSEELISWET 174 RASRAMIGHTDSAEAAPGTIRGDFSVHISRNVIHASDSVEGAQREIQLWFQSSELVSWAD 177 KTGRVMLGETNPADSKPGTIRGDFCIQVGRNIIHGSDSVESANKEIALWFKDEELVENKS 145 KTGRVMLGETNPADSKPGTIRGDLCIQVGRNIIHGSDSVDSANKEIALWFKDEELVEYKS 145 KTARLMIGETNPAHSLPGTIRGDFCIDVGRNVIHGSDSRESAEREIALWFQPDELVYWQD 161 * * * ******* ** ** *** * ** *** ** SHQ-SH--LCQV---- 183 GGQHSSIHPA------ 187 CAY-------EWYYEN 154 CAY-------EWYYEN 154 SAE-------SWIYE- 169 Sequence Figure 1 alignment of NM23-X4 Sequence alignment of NM23-X4. (A) Phylogenetic tree of human and Xenopus NM23 members. The tree was constructed using the Clustal W method with the parameter PAM250. (B) Nucleotide sequence alignment of NM23-X4 with NM23-H4, -X1, -X2, and -X3. Asterisks indicate identical amino acids among all NM23 family members. The previously reported important amino acid residues are shown as shaded boxes. Page 3 of 18 (page number not for citation purposes)
Neural Development 2009, 4:1 http://www.neuraldevelopment.com/content/4/1/1 Interaction Figure 2 between NM23 and CDKIs Interaction between NM23 and CDKIs. (A) Co-immunoprecipitation of NM23-X4 and p27Xic1 in COS7 cells. p27Xic1 interacts strongly with NM23-X4 in the presence of the proteasome inhibitor MG132. (B) Interaction of p27Xic1 with NM23- X1, -X2, -X3, -X4, -X5, or -X6. (C) Interaction of p27Xic1 with the human NM23-H1 and -H4 homologs. (D) NM23-X4 interacts strongly with human p21Cip1 and p57Kip2, but not with p27Kip1. All analyses were performed in COS7 cells in the presence of MG132 unless otherwise stated. Next, we wanted to know if p27Xic1 interacts with other NM23 family members. As shown in Figure 2B, NM23-X3 also binds strongly to p27Xic1, while all other NM23s tested interacted weakly. The human NM23-H1 and H4 interacted with p27Xic1 similarly to their Xenopus orthologs (Figure 2C). Taken together, these results suggest that NM23-X4 and/or X3 are likely to be biologically significant binding partners of p27Xic1. Mammals have three Cip/Kip CDKIs (p21Cip1, p27Kip1, and p57Kip2). Interestingly, NM23-X4 interacted with p21Cip1 and p57Kip2, but not with the human p27Kip1 (Figure 2D). We identified two other Xenopus CDKIs, p16Xic2 and p17Xic3, which also have gliogenic functions, similar to that of p27Xic1, although their expression levels in the developing retina are lower [30]. Interestingly, we found that p16Xic2 does not interact with NM23-X4 (unpublished data). Sequence alignment Page 4 of 18 (page number not for citation purposes)
Page 1 and 2: 5 January 2009 Neural Development w
Page 3: Neural Development 2009, 4:1 http:/
Page 7 and 8: Neural Development 2009, 4:1 http:/
Page 19: Neural Development 2009, 4:1 http:/

Neural Development - BioMed Central

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?