Crystal structures of Drosophila mutant translin and characterization of translin variants reveal the structural plasticity of translin proteins

被引:9
作者
Gupta, Gagan D. [1 ]
Makde, Ravindra D. [1 ]
Rao, Basuthkar J. [2 ]
Kumar, Vinay [1 ]
机构
[1] Bhabha Atom Res Ctr, High Pressure Phys Div, Bombay 400085, Maharashtra, India
[2] Tata Inst Fundamental Res, Dept Biol Sci, Bombay 400005, Maharashtra, India
关键词
chimeric translin; crystal structure; Drosophila melanogaster translin; oligomerization; P168S mutant;
D O I
10.1111/j.1742-4658.2008.06571.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Translin protein is highly conserved in eukaryotes. Human translin binds both ssDNA and RNA. Its nucleic acid binding site results from a combination of basic regions in the octameric structure. We report here the first biochemical characterization of wild-type Drosophila melanogaster (drosophila) translin and a chimeric translin, and present 3.5 angstrom resolution crystal structures of drosophila P168S mutant translin from two crystal forms. The wild-type drosophila translin most likely exists as an octamer/decamer, and binds to the ssDNA Bcl-CL1 sequence. In contrast, ssDNA binding-incompetent drosophila P168S mutant translin exists as a tetramer. The structures of the mutant translin are identical in both crystal forms, and their C-terminal residues are disordered. The chimeric protein, possessing two nucleic acid binding motifs of drosophila translin, the C-terminal residues of human translin, and serine at position 168, attains the octameric state and binds to ssDNA. The present studies suggest that the oligomeric status of translin critically influences the DNA binding properties of translin proteins.
引用
收藏
页码:4235 / 4249
页数:15
相关论文
共 40 条
[1]   Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].
Altschul, SF ;
Madden, TL ;
Schaffer, AA ;
Zhang, JH ;
Zhang, Z ;
Miller, W ;
Lipman, DJ .
NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402
[2]   The DNA binding activity of Translin is mediated by a basic region in the ring-shaped structure conserved in evolution [J].
Aoki, K ;
Suzuki, K ;
Ishida, R ;
Kasai, M .
FEBS LETTERS, 1999, 443 (03) :363-366
[3]   Isolation and characterization of a cDNA encoding a Translin-like protein, TRAX [J].
Aoki, K ;
Ishida, R ;
Kasai, M .
FEBS LETTERS, 1997, 401 (2-3) :109-112
[4]   A NOVEL GENE, TRANSLIN, ENCODES A RECOMBINATION HOTSPOT BINDING-PROTEIN ASSOCIATED WITH CHROMOSOMAL TRANSLOCATIONS [J].
AOKI, K ;
SUZUKI, K ;
SUGANO, T ;
TASAKA, T ;
NAKAHARA, K ;
KUGE, O ;
OMORI, A ;
KASAI, M .
NATURE GENETICS, 1995, 10 (02) :167-174
[5]   THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY [J].
BAILEY, S .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 :760-763
[6]   The Protein Data Bank [J].
Berman, HM ;
Westbrook, J ;
Feng, Z ;
Gilliland, G ;
Bhat, TN ;
Weissig, H ;
Shindyalov, IN ;
Bourne, PE .
NUCLEIC ACIDS RESEARCH, 2000, 28 (01) :235-242
[7]   Free R value: Cross-validation in crystallography [J].
Brunger, AT .
MACROMOLECULAR CRYSTALLOGRAPHY, PT B, 1997, 277 :366-396
[8]   Trax (Translin-associated factor X), a primarily cytoplasmic protein, inhibits the binding of TB-RBP (Translin) to RNA [J].
Chennathukuzhi, VM ;
Kurihara, Y ;
Bray, JD ;
Hecht, NB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (16) :13256-13263
[9]   Altering the GTP binding site of the DNA/RNA-binding protein, Translin/TB-RBP, decreases RNA binding and may create a dominant negative phenotype [J].
Chennathukuzhi, VM ;
Kurihara, Y ;
Bray, JD ;
Yang, JX ;
Hecht, NB .
NUCLEIC ACIDS RESEARCH, 2001, 29 (21) :4433-4440
[10]   Functional characterization of Drosophila Translin and Trax [J].
Claussen, Maike ;
Koch, Rafael ;
Jin, Zhao-Yang ;
Suter, Beat .
GENETICS, 2006, 174 (03) :1337-1347