Co‐expressed recombinant human Translin‐Trax complex binds DNA

Gupta, Gagan D.; Makde, Ravindra D.; Kamdar, Radhika Pankaj; D’Souza, Jacinta S.; Kulkarni, Meeta G.; Kumar, Vinay; Rao, B.S. Shankaranarayana

doi:10.1016/j.febslet.2005.05.007

Cited by 18 publications

(9 citation statements)

References 22 publications

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“…Translin selectivity towards RNA or DNA is also modulated in interactions with another translin-like protein known as trax. The trans-lin protein carries a functional nuclear export signal sequence, and the trax protein possesses a nuclear import signal sequence and is post-transcriptionaly stabilized by translin protein [7][8][9][10][11][12]. Translin and trax proteins are highly conserved in eukaryotes.…”

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confidence: 99%

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

et al. 2008

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Human translin, a 27-kDa protein, binds to ssDNA and RNA. It recognizes chromosomal breakpoint sequences at ssDNA ends with varying affinity [1], and is recruited to the nucleus concomitant with the induction of double-strand breaks by DNA-damaging agents and in many cases of lymphoid neoplasms that are robustly associated with T-cell receptor or immunoglobulin gene translocations [2,3]. The mouse translin has been proposed to function in regulating the expression of a variety of mRNA sequences by regulating RNA translocations and localization within and perhaps even across cells [4,5]. It has been proposed that GTP modulates the binding affinity of mouse translin towards RNA or DNA. For instance, its binding affinity towards RNA decreases by more than 50% but its DNA binding affinity remains essentially unaltered in the presence of GTP [6]. Translin selectivity towards RNA or DNA is also modulated in interactions with another translin-like protein known as trax. The translin protein carries a functional nuclear export signal sequence, and the trax protein possesses a nuclear import signal sequence and is post-transcriptionaly stabilized by translin protein [7][8][9][10][11][12]. Translin and trax proteins are highly conserved in eukaryotes. These interacting partners co-exist in vertebrates including human, mouse, chicken, Xenopus etc, in invertebrates such as Drosophila melanogaster (drosophila), and also in Schizosaccharomyces pombe fungus, and their complex has recently been suggested to regulate dendritic trafficking of RNAs [13].Two basic nucleic acid binding motifs (basic-1 and basic-2) have been identified from mutational studies on human and mouse translin [6,14]. It is surmised that the DNA-binding domain of human translin is formed by combination of its basic regions in a multimeric structure, and the loss of multimeric structure results in abrogation of its DNA binding abilities [14]. Currently, the crystal structures of human and mouse 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 Å 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 bind...

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confidence: 99%

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

et al. 2008

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“…Accordingly, it is thought to play a role in the subcellular transport and/or translational control of its target RNAs in these tissues (Han et al 1995a;Kobayashi et al 1998;Morales et al 1998;Muramatsu et al 1998;Wu and Hecht 2000;Yang et al 2003). Unlike Translin, Trax does not bind nucleic acids directly, but might be part of the RNA-or DNA-binding complex, thereby modulating the nucleic-acid-binding affinity of Translin (Chennathukuzhi et al 2001;Finkenstadt et al 2002;Gupta et al 2005).…”

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Functional Characterization of Drosophila Translin and Trax

Claußen¹,

Koch²,

Jin³

et al. 2006

Genetics

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The vertebrate RNA and ssDNA-binding protein Translin has been suggested to function in a variety of cellular processes, including DNA damage response, RNA transport, and translational control. The Translin-associated factor X (Trax) interacts with Translin, and Trax protein stability depends on the presence of Translin. To determine the function of the Drosophila Translin and Trax, we generated a translin null mutant and isolated a trax nonsense mutation. translin and trax single and double mutants are viable, fertile, and phenotypically normal. Meiotic recombination rates and chromosome segregation are also not affected in translin and trax mutants. In addition, we found no evidence for an increased sensitivity for DNA double-strand damage in embryos and developing larvae. Together with the lack of evidence for their involvement in DNA double-strand break checkpoints, this argues against a critical role for Translin and Trax in sensing or repairing such DNA damage. However, Drosophila translin is essential for stabilizing the Translin interaction partner Trax, a function that is surprisingly conserved throughout evolution. Conversely, trax is not essential for Translin stability as trax mutants exhibit normal levels of Translin protein.

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“…The gel shift complex was excised and analyzed for its composition on SDS-PAGE. Stoichiometrically, it was found that the minimum binding unit for ssDNA was a dimer of translin and monomer of trax which existed nearly in a ratio of 1:1, similar to that of a purified recombinant complex [56]. All these results, put together, suggested that heteromeric complex exhibits relatively more stable binding to ssDNA.…”

Section: Interactors Of Trax and Their Physiological Significancementioning

confidence: 75%