ΘΕΜΑ 12345 ΕΡΓΑΣΙA ΔΕΥΤΕΡΗ

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ΓΡΑΠΤΗ ΕΡΓΑΣΙΑ ΒΙΟΛΟΓΙΑΣ ΕΑΠ

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<p> 1 , (2--D- D- DNA RNA ) . . . DNA RNA , ATP, GTP, CTP UTP. , de novo . de novo , , . To . , , (UMP) (CMP). , , C . . , , . -, NH3 CO2. (PP) , . </p> <p> , HPLC Mass Spectrometry. . PP , Lesch-Nyhan , . . ( ), ( ). , RNA, . [1]</p> <p> 1.1 .</p> <p> 1.2. 1.1. , DNA, -. [2]</p> <p>2</p> <p> (A) (G) (C) (U) (T)).</p> <p> (AMP) (GMP) (CMP) (UMP) (TMP)</p> <p> 1.1 RNA ( </p> <p> 1.2 . </p> <p> - . [2]</p> <p> , . , (UMP), RNA. [2]</p> <p> (CMP) , - (UDP) - (UTP), ATP. [2] UMP + ATP UDP + ADP UDP + ATP UTP + ADP - (CTP) , . [2] UTP + (3) + ATP CTP + ADP + Pi + </p> <p>4</p> <p> 1.3 de novo </p> <p> - -, , - . [2]</p> <p> , . . [2]</p> <p>6</p> <p> 1.4 .</p> <p> C , 1.5. [2] , . [2]</p> <p> 1.5 C .</p> <p> . . . ( ), , . , 3 CO2. [2]</p> <p> 1.6 .</p> <p>8</p> <p> 1.7 AMP . . pH . , .</p> <p> -, 3 CO2. , . - , -, 3 CO2. , -. [2]</p> <p> 1.8 . </p> <p> (PP) reviews . - , . PP , . , . . (high-performance liquid chromatography, HPLC), (mass spectrometry), , . . , , . [4] , . , . </p> <p>10</p> <p> PP . . PP , . o . . [5] , , , , , , </p> <p> , , , , gout . , (.. 5-fluorouracil azathioprine ). , . [5]</p> <p> (gouty arthritis) , . ( + = ), </p> <p> . [3] </p> <p> , . 1/3 , 2/3 . 3,5-7,2 mg/dL , 2,6-6,0 mg/dL 2,0-5,5 mg/dL . 7 mg/dL (, hyperuricaemia), , , ( ), , . , (). , . . 40 , . o , . : (peak), , . , , . . , 30% . [3] </p> <p> ( Gout = , ipperlein), , , </p> <p>12</p> <p> . Gout , . . , . H ( Gouty rthritis), , , , (, , , ). ( ), , , </p> <p> , , ( , ). () , . [3]</p> <p> (). , () . :</p> <p>1. , , , , .. 2. . 3. , , . 4. , (.. Lesch-Nyhan). 5. ( ). 6. : . </p> <p>7. . 8. pH . 9. (, , ). , , ( ). , , . [3]</p> <p> ( Lesh-Nyhan) GPRT (----) . H : () . () Lesch-Nyhan ( ). () (Glycogen Storage Disease). () . , (). , . [3]</p> <p> :</p> <p> (Purine turnover): , , . , , . . X .</p> <p>14</p> <p> (Cytotoxic drugs). , , , . : ( </p> <p>, ). . [3] , . , . [3]</p> <p> Lesch-Nyhan</p> <p> NCBI, Lesch-Nyhan (LNS) : , , , , , . (van der Zee et al., 1968). [6] (HPRT) Lesch-Nyhan Kelley-Seegmiller. To LNS . , KelleySeegmiller . , KelleySeegmiller, LNS. , Kelley-Seegmiller (gout). (1,5 8%), </p> <p> (Jinnah and Friedman, 2001). [7] 1979, Bakay HPRT, , , . ( ). HPRT , . [8] Page , HPRT, . 7,5% . . , , . [9] 2008, Hladnik , HPRT . Lesch-Nyhan, , , . , , , . , gout IQ. , . [10] 1977, Vogel Lesch-Nyhan, . [11] 1965 (Hoefnagel et al.) HPRT. [12]</p> <p>16</p> <p>1. McMurry J. , , . 29, . 1395-1398, 2004. 2. . ,, .; . 6, . 134-158, 2000. 3. . 4. Jurecka A. Inborn errors of purine and pyrimidine metabolism, J Inherit Metab Dis. 2009 Apr; 32(2):247-63, Epub 2009 Mar 15. 5. Simmons HA, Duley JA, FairbanksLD, McBride MB. When to investigate for pyurine and pyrimidine disorders. Introduction and review of clinical and laboratory indications., J Inherit Metab Dis. 1997 Jun; 20(2):214-26. 6. Van der Zee, S. P. M.; Schretlen, E. D. A. M.; Monnens, L. A. H. : Megaloblastic anaemia in the Lesch-Nyhan syndrome. (Letter) Lancet 291: 1427 only, 1968. Note: Originally Volume I. 7. Jinnah, H. A.; Friedmann, T. : Lesch-Nyhan disease and its variants.In: Scriver, C. R.; Beaudet, A. L.; Sly, W. S.; Valle, D. (eds.) : The Metabolic &amp; Molecular Bases of Inherited Disease. Vol. II. 8th ed. New York: McGrawHill 2001. P. 2537. 8. Bakay, B.; Nissinen, E.; Sweetman, L.; Francke, U.; Nyhan, W. L. : Utilization of purines by an HPRT variant in an intelligent, nonmutilative patient with features of the Lesch-Nyhan syndrome. Pediat. Res. 13: 13651370, 1979. PubMed ID : 523196 9. Page, T.; Nyhan, W. L.; Morena de Vega, V. : Syndrome of mild mental retardation, spastic gait, and skeletal malformations in a family with partial deficiency of hypoxanthine-guanine phosphoribosyltransferase. Pediatrics 79: 713-717, 1987. PubMed ID : 3575027 10. Hladnik, U.; Nyhan, W. L.; Bertelli, M. : Variable expression of HPRT deficiency in 5 members of a family with the same mutation. Arch. Neurol. 65: 1240-1243, 2008. PubMed ID : 18779430 11. Vogel, F. : A probable sex difference in some mutation rates. (Editorial) Am. J. Hum. Genet. 29: 312-319, 1977. PubMed ID : 868880 12. Hoefnagel, D.; Andrew, E. D.; Mireault, N. G.; Berndt, W. O. : Hereditary choreoathetosis, self-mutilation and hyperuricemia in young males. New Eng. J. Med. 273: 130-135, 1965. PubMed ID : 14303658 </p> <p> 2 , RNA . RNA , , , . RNA . RNA -P, , , HDV glmS. . , , . RNA , . , . , </p> <p>18</p> <p> . H . . RNA . , , . , . . HDV. glmS . RNA o RNA , .. . Cech RNA . ( 413 ) Tetrahymena thermophila. , Zaug . Thermophila in trans, (.. RNA). Foster Symons RNA (virusoid RNAs) RNA . Uhlenbeck , (hammerhead ribozyme), . RNA </p> <p>,</p> <p>-</p> <p>RNA,</p> <p> . Haseloff Gerlach , in trans. , in vivo. in vivo 60% . , . [1] in vivo , in vitro . . [1] MichaelisMenton. , Kcat/Km. 108 -1 min-1. cat/Km , Km Kcat , , [1]. , , . , (single turnover reaction), . (Kalavrizioti et al., 2006). [6] in vitro, : gel , </p> <p>20</p> <p> . . , . : , . , . RNA. , . [1] . , . [1] RA . (self-splicing RNA) RNA P (RNase P) tRNAs, RNA . 1980, RNA (small nucleolytic ribozymes or RNA-based enzymes). : (hammerhead) , (hairpin ribozyme), (hepatitis delta virus, HDV) Neurospora crassa Varkud satellite (VS). RNA RNA (ligation reaction). [2] , , 3,5- RNA. , RNA RNase P tRNAs ( 1a). , o (transesterification), </p> <p> ( 1b) ( 1c,d). 3,5- , RNA 2,3- 5 ( 1b). , o , pH . RNA, . in vitro leadzyme, RNase A riboswitch glmS, RNA . [2] , ( 1c,d). 3-OH 5 , , 3 . 3- . ( 1c). , 2,3 , 2,5- . , 2 ( 1d). RNA , . tRNA Eukarya Archaea , , 2,3- . [2]</p> <p>22</p> <p> 2.1 RNA. (a) tRNA RNase P 5- tRNA 3- 5 . (b-d) (b) , (c) , (d) . (b), 2- 3,5- , 2,3- 5-. </p> <p> (c), 3- (G) 5- 5 , 5 . , 5 3,5 3 , . (d), , . (e) .</p> <p> RNA . RNA , RNA ( 1e). RNA . RNA rRNA. [2] , RNA , riboswitch . 1 RNAs. RNA ; , RNA, . [2]</p> <p>24</p> <p> 2.1 riboswitches.</p> <p> RNA , . , </p> <p>. , , . , , . [2] , . , - 7 - . , ( 2.2). . , , , . [2]</p> <p> 2.2 . (a) </p> <p>26</p> <p> RNA . (b) RNA . , ( ) .</p> <p> , in silico . , RNA , , , . , , , . , (non-Watson-Crick base pairing). , ( 2.3b). non-Watson-Crick . , Watson-Crick , HDV ( 2.3a). . [2]</p> <p> 2.3 . (a) HDV (b) . To HDV (convoluted pseudoknotted topology). , Watson-Crick ( ), Hoogsteen ( ) ( ). (cis , ) (trans-, ). , . syn- Bold. .</p> <p>, in vitro, , , . in vitro . </p> <p>28</p> <p>. , in vitro , . [2] . ; DNA ; , , . DNA DNA; [3] RNA , , . DNA RNA . [3] RNA , DNA . Walter Gilbert RNA . RNA . DNA RNA RNA , RNA. , RNA . [4]</p> <p> RNA </p> <p> ( 2.4). </p> <p> , 5 5 . , 5 3 . . . Steitz Steitz . . [5] X. 2004. , RN, , . , , , . . [5]</p> <p>30</p> <p> 2.4 . . U-1 O 3. G O3. .</p> <p> HDV RNA . . . , RNA. pKa 6,8. pH. [5] </p> <p> HDV - RNA . 2 5 RNA, 2,3- 5- ( 2.5). . HDV , 5- 3 C75, . C75 O5 . [5]</p> <p> 2.5 RNA HDV.</p> <p>32</p> <p> tRNAs, -tRNA tRNA P. To P 3 O RNA, A76. H P -. S. [5] 76 2- P . . , . rRNA 2- 2451 . [5] , . pa 12, 2- , pH. [5] 76 2- - 3- ( 2.6). 3 , - 76 3 76 2-. 2-. . [5]</p> <p> 2.6 . .</p> <p> GlmS RNA , ( 2.7). glmS riboswitch 5 glycosamine-6phosphate (GlcN6P) synthetase. GlcN6P mRNA, . GlcN6P , in vitro. H </p> <p>34</p> <p> . GlcN6P . [5] GlcN6P ; 2 . C2 , . 5 . [5]</p> <p> 2.7 RNA glmS GlcN6P. . G33 , .</p> <p> , ( ). , , : ; DNA, ; , DNA; , . , , , , . ; ; </p> <p>, .. . ; . . . , : 36</p> <p> ; DNA. .</p> <p> 1. RNA : rRA, mRNA tRNA. ; 2. RNA (ncRNA) RNA . . 4 5 RNA. 3. RNA. ; . 4. RNAs . 2. 5. . , , , Neurospora crassa . ~35 ~164 . , 200 1500 , 4 . . 300 3000 . , RNA </p> <p> P (RNase P) . ~ 400-500 . 1. 6. . - Mg2+ - . . . 7. . 3 . 8. . , . , , . 9. 1990, Noller - . . - ( ) RNA. . . . , , .</p> <p>38</p> <p> RNA RNase P</p> <p> Neurospora crassa</p> <p> 1</p> <p> 2</p> <p>40</p> <p> 3 </p> <p> RNA: </p> <p> 4</p> <p>42</p> <p>Obj100</p> <p> 5 1. Antony M. Jose Ribozyme therapy: RNA Enzymes to the Resque, Yale Journal of Biology and Medicine 75 (2002), pp 215-219. 2. Cristian Hamman and Eric Westhof Searcing genomes for ribozymes and riboswitches, Genome Biology 2007, 8:210. 3. , , , , 33. 4. Lubert Streyer , , . 222-223. 5. Scott Strobel and Jesse C. Cochrane RNA Catalysis: Ribozymes, Ribosomes and Riboswitches Curr Opin Chem Biol. 2007 December; 11(6): 636-643. 6. Kalavrizioti D., Vourekas A., Stamatopoulou V., Toumpeki C., Giannouli S., Stathopoulos K., Drainas D., RNA Mediated Therapeutics: From Gene Inactivation to Clinical Application Current Topics in Medicinal Chemistry, Volume 6, Number 16, p. 1737-1758, 2006. 3 , , . , . 1, 2, 3, 4, 5, 6 7. . 1 1975 . </p> <p>Dejerine-Sottas</p> <p>. , , Charot-Marie-Tooth 1. PMP22 2 5% . Li-Fraumeni (LFS1, LFS2 LFS3) Li-Fraumeni-like. 1994 (Birch et al.) . LFS . (FGFR2), Pfeiffer Crouzon. Pfeiffer 1 . , 342 , Crouzon. , Denys-Drash WT1 , Wilms.</p> <p> . , , (rearrengments)...</p>