Dissertation > Excellent graduate degree dissertation topics show

Characterization of RadA Homologues from the Hyperthermophilic Archaeon Sulfolobus Tokodaii

Author: ZhuShanShan
Tutor: ShenYuLong
School: Shandong University
Course: Microbiology
Keywords: Hyperthermophilic Crenarchaeota Homologous recombination RadA homologous protein RadA
CLC: Q93
Type: Master's thesis
Year: 2008
Downloads: 45
Quote: 0
Read: Download Dissertation

Abstract


Homologous recombination DNA repair pathway is common in the biological, it has an important role to maintain the stability of the genome for the repair of DNA breaks, to restart replication fork and eukaryotic meiosis. The archaea homologous recombination only in its infancy, however, found that a protein similar to eukaryotic homologous recombination mechanism: restructuring early breaking double-stranded protein Mrell and Rad50 are; wide archaea single-chain binding protein RPA; the recombinase in archaea (recombinase) RadA, which has about 40% homology to the Rad51 eukaryotes, only about 20% homology with RecA; similar to true The presence of the the Rad51 homologous proteins, nuclear biological RadA homologous proteins also exist in a wide archaea. RecA/Rad51/RadA key protein homologous recombination in the three domains of biological promote homologous recombination strand exchange to form a Holliday junction structure. Participation of many proteins in bacteria and eukaryotes mediated RecA/Rad51 form a nucleoprotein filament or strand exchange, the bacteria RecBCD RecFOR complex eukaryotes Rad52, Rad54, Rad55 and Rad57 etc.. The presence of the Rad51 homologous proteins including the yeast Rad55/Rad57 complex, in higher eukaryotes Rad51B-Rad51C-Rad51D-of XRCC2 and Rad51C-XRCC3 complex in eukaryotes. Genetic analysis showed that they are involved in homologous recombination, which Rad55/Rad57 stable Rad51 nucleoprotein filament structure has been identified. The homologous proteins wide archaea RadA RadB. However, there are not the Crenarchaeota RadA homologous proteins of reports. Homologous proteins in to find four RadA thermophilic spring Archaea Sulfolobus tokodaii Str.7 genome sequence analysis, phylogenetic analysis showed that they are more similar to eukaryotes Rad55, and thus we named stRad55A ( ST0579), stRad55B (ST0838), stRad55C (ST1830) and stRad55D (ST2522). Using RT-PCR analysis of the S.tokodaii radA, rad55A, rad55B rad55C and rad55D gene transcription induced by ultraviolet radiation, found the radA, rad55A and rad55B gene transcription is induced by ultraviolet, suggesting that these proteins may be involved in DNA repair. These protein and S.tokodaii of single-stranded binding protein SSB, which, RadA, stRad55A stRad55C and SSB expression in E.coli bacteria that can be expressed in soluble form, stRad55B and stRad55D form of inclusion bodies in E.coli expressed. RadA, stRad55A and stRad55C molecular sieve chromatography The results show that in solution exist in the form of oligomers, respectively, SSB exist in monomeric form. Further biochemical characterization of the the stRad55A, stRad55B and stRad55C found that (1) stRad55A have a lot of similar nature in Rad52: relative to the double-stranded DNA (dsDNA), single-stranded DNA (ssDNA) has a greater affinity; ssDNA-dependent ATPase activity; can lift SSB RadA chain exchange activity; mediated RadA ssDNA chain bonded to the SSB covered; have interaction with RadA and SSB. However, there are with the the Rad52 different nature: the inhibition of the activity of the RadA; not detected in the strand exchange reaction system without the SSB to its DNA annealing activity. (2) co-expression through stRad55B with RadA give stRad55B soluble form of the protein and purified stRad55B protein, having an interactable Notes stRad55B with RadA, can form a composite body; relative the of dsDNA, stRad55B on ssDNA having a greater affinity ; having weak ATPase activity does not depend on the DNA; inhibit RadA activity of strand exchange in the reaction system without the SSB. The current nature of the biochemical data show the nature of stRad55B more similar to RadB. (3) stRad55C stRad55A more binding ssDNA, ssDNA-dependent ATPase activity; the in stRad55C with S.tokodaii RadA and Hjc have interaction; SSB strand exchange reaction system inhibit RadA activity; promote Hjc cutting the activity of the Holliday junction. We S.tokodaii study is the first time the Crenarchaeota RadA homologous proteins were identified. The data obtained can not completely explain the specific role of the protein in the cell, but some of the identification of biochemical properties as well as protein-protein interaction detection result confirmed that these proteins are involved in DNA repair. The specific mechanism of action of the protein in the cells requires the identification and genetic analysis of biochemical properties.

Related Dissertations

  1. Select of Biofilm Formation Mutants and Cloning and Primary Function Analysis of tatC Gene from Pseudomonas Fluorescens Strain 7-14,S432.4
  2. Implantation of Mesenchymal Stem Cells (MSCs) Carried by LINK-N/RADA-16 to the Intervertebral Disc : A Study to Repair the Disc,R687.3
  3. Modification of Phosphoenolpyruvate: Carbohydrate Phosphotransferase System of Escherichia Coli by Genetic Engineering and Characterization of Mutants,Q78
  4. Increase of the Yield of γ-decalactone by Genetically Modified Strain of Yarrowia Lipolytica,TS202.3
  5. The Effect of Astaxanthin Yield by Overexpressing GGPP Synthase in Phaffia Yeast,TQ929
  6. Lidamycin a cloning the bacteria atrA homologous genes and its function,TQ465
  7. Construction of Mouse HSP10 SiRNA and Overexpression Adenovirus Vector and Investigation of Its Role on the Apoptosis of Mouse Ovarian Granulosa Cells,R363
  8. Study on ALD4 Gene Disruption and GPD1 Silencing in Saccharomyces Cerevisiae Industrial Strain Y01,Q78
  9. Construction of Recombinant Adenovirus Vector Co-expressing P12A and 3C Genes of FMDV O Virus,S852.65
  10. A method of oligonucleotide-mediated E. coli gene knockout or point mutations in the,Q78
  11. Construction and Functional Analysis of the Common Allelic Gene Double-knockout Vector System,Q78
  12. Construction Expression Cellulase Recombinant Strain of Lactobacillus Reuteri Without Resistance Marker,S816
  13. Prestin Gene-Related Human Deafness Disease Animal Model is Established,R-332
  14. Construction and Expression of Marker-free Targeting Vector of Human Leukemia Inhibitory Factor,R733.7
  15. The Construction of BAC-HSV-1 Strain HF with GFP Reporter Gene and the Research of Its Infectious Progeny Virus,R373
  16. Relationship between the Mutation of KatG in Mycobacterium Tuberculosis and Drug-resistance of INH,R52
  17. Infectivity Analysis of AcMNPV Deleted pk-1 Gene,Q78
  18. Clonging and Functional Identification of the Genes luxS and pilA Related To Biofilm Formation of Haemophilus Parasuis,S852.61
  19. Construction of Recombinant Adenovirus of VP2 Protein of Infectious Bursal Disease Virus JS Strain and Biological Characterization of the Recombinant Virus,S852.65
  20. Study on Irp1 and Irp2 Genes of HPI in Escherichia Coil,S852.61
  21. Construction of SEF14 Fimbrial Deletion Mutants from Salmonella Enteritidis and Analysis of the Related Functions,S852.61

CLC: > Biological Sciences > Microbiology
© 2012 www.DissertationTopic.Net  Mobile