DLGAP5
Disksovski velikopridruženi protein 5 (DAP-5), znan i kao diskovski veliki homolog 7 (DLG7) ili hepatomski nadregulirani protein (HURP), je protein koji je kod ljudi kodiran genom DLGAP5.[5][6][7][8]
Pomoći PCR analize hibridnog panela ćelija čovjek-glodar, Nomura et al. (1994) locirali su gen DLGAP5 na hromosomu 14.[9] Primjenom fluorescentne hibridizacije in situ, Bassal et al. (2001) precirirali su mapiranje gena DLGAP5 na hromosomskoj poziciji 14q22.2-q23.2.[10] I Hartz (2018) mapirala je gen DLGAP5 na hromosomskoj lokaciji 14q22.3, na osnovu poravnavanja sekvence DLGAP5 (GenBank D13633) sa genomskom sekvencom (GRCh38).[11]
Tsou et al (2003) utvrdili su da gen DLGAP5 ima 19 egzona i da se proteže približno na 43,3 kb. Egzon 1 je nekodirajući.[12] DLG7 je kinetohorni protein koji stabilizira mikrotubule u blizini hromosoma. DLG7 kontrolira dinamiku vretenskog aparatae, pospješujući interkinetohornu napetost i efikasno hvatanje kinetohora.[13] DGL7 je dio Ran-ovisnog kompleksa.[14] Stabilizacija DGL7 u ćeliji nastaje uslijed fosforilacije aurora A-kinaza.[15] Ekspresija DGL7 nađena je u karcinomu[16] i matičnim ćelijama.[17]
Aminokiselinska sekvenca uredi
Dužina polipeptidnog lanca je 846 aminokiselina, a molekulska težina 95.115 Da.[18]
| 10 | 20 | 30 | 40 | 50 | ||||
|---|---|---|---|---|---|---|---|---|
| MSSSHFASRH | RKDISTEMIR | TKIAHRKSLS | QKENRHKEYE | RNRHFGLKDV | ||||
| NIPTLEGRIL | VELDETSQGL | VPEKTNVKPR | AMKTILGDQR | KQMLQKYKEE | ||||
| KQLQKLKEQR | EKAKRGIFKV | GRYRPDMPCF | LLSNQNAVKA | EPKKAIPSSV | ||||
| RITRSKAKDQ | MEQTKIDNES | DVRAIRPGPR | QTSEKKVSDK | EKKVVQPVMP | ||||
| TSLRMTRSAT | QAAKQVPRTV | SSTTARKPVT | RAANENEPEG | KVPSKGRPAK | ||||
| NVETKPDKGI | SCKVDSEENT | LNSQTNATSG | MNPDGVLSKM | ENLPEINTAK | ||||
| IKGKNSFAPK | DFMFQPLDGL | KTYQVTPMTP | RSANAFLTPS | YTWTPLKTEV | ||||
| DESQATKEIL | AQKCKTYSTK | TIQQDSNKLP | CPLGPLTVWH | EEHVLNKNEA | ||||
| TTKNLNGLPI | KEVPSLERNE | GRIAQPHHGV | PYFRNILQSE | TEKLTSHCFE | ||||
| WDRKLELDIP | DDAKDLIRTA | VGQTRLLMKE | RFKQFEGLVD | DCEYKRGIKE | ||||
| TTCTDLDGFW | DMVSFQIEDV | IHKFNNLIKL | EESGWQVNNN | MNHNMNKNVF | ||||
| RKKVVSGIAS | KPKQDDAGRI | AARNRLAAIK | NAMRERIRQE | ECAETAVSVI | ||||
| PKEVDKIVFD | AGFFRVESPV | KLFSGLSVSS | EGPSQRLGTP | KSVNKAVSQS | ||||
| RNEMGIPQQT | TSPENAGPQN | TKSEHVKKTL | FLSIPESRSS | IEDAQCPGLP | ||||
| DLIEENHVVN | KTDLKVDCLS | SERMSLPLLA | GGVADDINTN | KKEGISDVVE | ||||
| GMELNSSITS | QDVLMSSPEK | NTASQNSILE | EGETKISQSE | LFDNKSLTTE | ||||
| CHLLDSPGLN | CSNPFTQLER | RHQEHARHIS | FGGNLITFSP | LQPGEF |
- Simboli
Kloniranje i ekspresija uredi
DLGAP5 je protein povezan s mitotskim mikrotubulama koji pojačava njihov u polimerizaciju i promovira pravovremeno stvaranje i stabilnost bipolarnog vretena .
Sekvenciranjem klonova dobijenih iz frakcionirane veličine nezrelih mijeloidnih ćelijskih linija KG-1, Nomura et al. (1994) klonirali DLGAP5, koji su označili KIAA0008. Izvedeni protein sadrži 805 aminokiselina. Northern blot analiza otkrila je visoku ekspresiju KIAA0008 u timusu, sjemenicima i debelom crijevu, te umjerenu ekspresiju u placenti, bubrezima, jajnicima i tankom crijevu. Ekspresija je bila niska u leukocitima periferne krvi, a nije je bilo u srcu, mozgu, plućima, jetri, skeletnim mišićima, gušterači i prostati. Ekspresija KIAA0008 bila je visoka u KG-1 i u HeLa ćelijama. Koristeći Northern blot analizu, Bassal et al. (2001) otkrili su visoku ekspresiju KIAA0008 u timusu, sjemenicima i debelom crijevu. U HeLa ćelijama, ekspresija KIAA0008 inducirana je tokom S faze ćelijskog ciklusa i održavana je tokom G2 i M-faze.
Tsou et al. (2003) klonirali su DLGAP5, koji su nazvali HURP, iz humane Hep3B cDNA biblioteke hepatoma. Izvedeni protein od 846 aminokiselina ima navodni nuklearni eksportni signal bogat leucinom, dvije kutije za uništavanje, KEN-kutiju i domen sa upredenom zavojnicom. Također ima približno 100-aminokiselinski gvanilat-kinaze-pridruženi protein (GKAP, ili DLGAP1; 605445) domen homologije-1. HURP dijeli 61% aminokiselinskog identiteta s utvrđenim proteinom miša Hurp od 808 aminokiselina. Kvantitativna RT-PCR analiza ljudskih tkiva otkrila je obilnu ekspresiju HURP-a u fetusnoj jetri, pa koštanom srži, sjemenicima, debelom crijevu i placenti. Ekspresija HURP-a bila je niska na tranziciji G1/S u sinhroniziranim HeLa ćelijama, porasla je kroz S fazu i G2/M, a vrhunac je dosegla u vrijeme kada su ćelije izašle iz mitoza. Imunofluorescentnom analizom otkriven je endogeni HURP HeLa ćelija u citoplazmi i jedru, tokom interfaze, ali je lokaliziran na vretenasta vlakna u mitotskim ćelijama.
Funkcija gena uredi
Tsou et al. (2003) otkrili su da je ekspresija Hurpa indukovana tokom regeneracije mišje jetre nakon parcijalne hepatektomije. Prekomerna ekspresija HURP-a u 293T ćelijama promovirala je preživljavanje ćelija u niskom serumskom i povišenom ćelijskom potencijalu za rast, neovisan o sidrištu.
Koristeći genomski zaslon, Wong i Fang (2006) identificirali su HURP kao mitotski regulator. U poređenju sa kontrolama, vretenske mikrotubule bez HURP-a bile su manje stabilne, sporije polimerizirane i pokazivale veće stope prometa. HURP-osiromašene ćelije imale su nevezane kinetohore na neporavnatim hromosomima i kinetohore pod djelomičnom napetošću na poravnatim. Loše poravnanje hromozoma duž metafazne ploče u ćelijama osiromašenim HURP-om rezultiralo je aktivacijom vretena i odloženim početkom anafaze; međutim, zadržavanje kontrolne tačke nije održano i ćelije su pokrenule anafazu u prisustvu djelimične napetosti kinetohora i neporavnatih hromosoma. Prekomjerna ekspresija HURP-a u HeLa ćelijama rezultirala je hiperstabilizacijom mitotskog vretena i smanjenom napetošću na sestrinskim kinetohorima.[19]
Reference uredi
- ^ a b c GRCh38: Ensembl release 89: ENSG00000126787 - Ensembl, maj 2017
- ^ a b c GRCm38: Ensembl release 89: ENSMUSG00000037544 - Ensembl, maj 2017
- ^ "Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
- ^ "Mouse PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
- ^ Nomura N, Miyajima N, Sazuka T, Tanaka A, Kawarabayasi Y, Sato S, Nagase T, Seki N, Ishikawa K, Tabata S (Dec 1995). "Prediction of the coding sequences of unidentified human genes. I. The coding sequences of 40 new genes (KIAA0001-KIAA0040) deduced by analysis of randomly sampled cDNA clones from human immature myeloid cell line KG-1". DNA Res. 1 (1): 27–35. doi:10.1093/dnares/1.1.27. PMID 7584026.
- ^ Nomura N, Miyajima N, Sazuka T, Tanaka A, Kawarabayasi Y, Sato S, Nagase T, Seki N, Ishikawa K, Tabata S (Dec 1995). "Prediction of the coding sequences of unidentified human genes. I. The coding sequences of 40 new genes (KIAA0001-KIAA0040) deduced by analysis of randomly sampled cDNA clones from human immature myeloid cell line KG-1 (supplement)". DNA Res. 1 (1): 47–56. doi:10.1093/dnares/1.1.47. PMID 7584028.
- ^ "Entrez Gene: DLG7 discs, large homolog 7 (Drosophila)".
- ^ Bassal S, Nomura N, Venter D, Brand K, McKay MJ, van der Spek PJ (septembar 2001). "Characterization of a novel human cell-cycle-regulated homologue of Drosophila dlg1". Genomics. 77 (1–2): 5–7. doi:10.1006/geno.2001.6570. PMID 11543626.
- ^ Nomura, N., Miyajima, N., Sazuka, T., Tanaka, A., Kawarabayasi, Y., Sato, S., Nagase, T., Seki, N., Ishikawa, K., Tabata, S. Prediction of the coding sequences of unidentified human genes. I. The coding sequences of 40 new genes (KIAA0001-KIAA0040) deduced by analysis of randomly samples cDNA clones from human immature myeloid cell line KG-1. DNA Res. 1: 27-35, 1994. Note: Erratum: DNA Res. 2: 210 only, 1995. PubMed: 7584026
- ^ Bassal, S., Nomura, N., Venter, D., Brand, K., McKay, M. J., van der Spek, P. J. Characterization of a novel human cell-cycle-regulated homologue of Drosophila dlg1. Genomics 77: 5-7, 2001. PubMed: 11543626
- ^ Hartz, P. A. Personal Communication. Baltimore, Md. 1/31/2018.
- ^ Tsou, A.-P., Yang, C.-W., Huang, C.-Y. F., Yu, R. C.-T., Lee, Y.-C. G., Chang, C.-W., chen, B.-R., Chung, Y.-F., Fann, M.-J., Chi, C.-W., Chiu, J.-H., Chou, C.-K. Identification of a novel cell cycle regulated gene, HURP, overexpressed in human hepatocellular carcinoma. Oncogene 22: 298-307, 2003. PubMed: 12527899
- ^ Wilde A (juni 2006). ""HURP on" we're off to the kinetochore!". J. Cell Biol. 173 (6): 829–31. doi:10.1083/jcb.200605150. PMC 2063907. PMID 16785318.
- ^ Koffa MD, Casanova CM, Santarella R, Köcher T, Wilm M, Mattaj IW (april 2006). "HURP is part of a Ran-dependent complex involved in spindle formation". Curr. Biol. 16 (8): 743–54. doi:10.1016/j.cub.2006.03.056. PMID 16631581.
- ^ Yu CT, Hsu JM, Lee YC, Tsou AP, Chou CK, Huang CY (juli 2005). "Phosphorylation and stabilization of HURP by Aurora-A: implication of HURP as a transforming target of Aurora-A". Mol. Cell. Biol. 25 (14): 5789–800. doi:10.1128/MCB.25.14.5789-5800.2005. PMC 1168800. PMID 15987997.
- ^ Sanderson HS, Clarke PR (juni 2006). "Cell biology: Ran, mitosis and the cancer connection". Curr. Biol. 16 (12): R466–8. doi:10.1016/j.cub.2006.05.032. PMID 16782004.
- ^ Gudmundsson KO, Thorsteinsson L, Sigurjonsson OE, et al. (juni 2007). "Gene expression analysis of hematopoietic progenitor cells identifies Dlg7 as a potential stem cell gene". Stem Cells. 25 (6): 1498–506. doi:10.1634/stemcells.2005-0479. PMID 17322106.
- ^ "UniProt, Q15398". Pristupljeno 23. 7. 2021.
- ^ Wong, J., Fang, G. HURP controls spindle dynamics to promote proper interkinetochore tension and efficient kinetochore capture. J. Cell Biol. 173: 879-891, 2006. PubMed: 16769820
Dopunska literatura uredi
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- Bassal S, Nomura N, Venter D, et al. (2001). "Characterization of a novel human cell-cycle-regulated homologue of Drosophila dlg1". Genomics. 77 (1–2): 5–7. doi:10.1006/geno.2001.6570. PMID 11543626.
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- Tsou AP, Yang CW, Huang CY, et al. (2003). "Identification of a novel cell cycle regulated gene, HURP, overexpressed in human hepatocellular carcinoma". Oncogene. 22 (2): 298–307. doi:10.1038/sj.onc.1206129. PMID 12527899.
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- Hsu JM, Lee YC, Yu CT, Huang CY (2004). "Fbx7 functions in the SCF complex regulating Cdk1-cyclin B-phosphorylated hepatoma up-regulated protein (HURP) proteolysis by a proline-rich region". J. Biol. Chem. 279 (31): 32592–602. doi:10.1074/jbc.M404950200. PMID 15145941.
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- Yu CT, Hsu JM, Lee YC, et al. (2005). "Phosphorylation and stabilization of HURP by Aurora-A: implication of HURP as a transforming target of Aurora-A". Mol. Cell. Biol. 25 (14): 5789–800. doi:10.1128/MCB.25.14.5789-5800.2005. PMC 1168800. PMID 15987997.
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- Wong J, Fang G (2006). "HURP controls spindle dynamics to promote proper interkinetochore tension and efficient kinetochore capture". J. Cell Biol. 173 (6): 879–91. doi:10.1083/jcb.200511132. PMC 2063914. PMID 16769820.
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Vanjski linkovi uredi
- DLG7 protein, human na US National Library of Medicine Medical Subject Headings (MeSH)