Functional specialization of Chlamydomonas reinhardtii cytosolic thioredoxin h1 in the response to alkylation-induced DNA damage

Nandita Sarkar, Stéphane Lemaire, Danxia Wu-Scharf, Emmanuelle Issakidis-Bourguet, Heriberto Cerutti

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

DNA damage occurs as a by-product of intrinsic cellular processes, like DNA replication, or as a consequence of exposure to genotoxic agents. Organisms have evolved multiple mechanisms to avoid, tolerate, or repair DNA lesions. To gain insight into these processes, we have isolated mutants hypersensitive to DNA-damaging agents in the green alga Chlamydomonas reinhardtii. One mutant, Ble-1, showed decreased survival when it was treated with methyl methanesulfonate (MMS), bleomycin, or hydrogen peroxide (H2O 2) but behaved like the wild type when it was exposed to UVC irradiation. Ble-1 carries an extensive chromosomal deletion that includes the gene encoding cytosolic thioredoxin h1 (Trxh1). Transformation of Ble-1 with a wild-type copy of Trxh1 fully corrected the MMS hypersensitivity and partly restored the tolerance to bleomycin. Trxh1 also complemented a defect in the repair of MMS-induced DNA strand breaks and alkali-labile sites. In addition, a Trxh1-β-glucuronidase fusion protein translocated to the nucleus in response to treatment with MMS. However, somewhat surprisingly, Trxh1 failed to correct the Ble-1 hypersensitivity to H2O2. Moreover, Trxh1 suppression by RNA interference in a wild-type strain resulted in enhanced sensitivity to MMS and DNA repair defects but no increased cytotoxicity to H2O2. Thioredoxins have been implicated in oxidative-stress responses in many organisms. Yet our results indicate a specific role of Chlamydomonas Trxh1 in the repair of MMS-induced DNA damage, whereas it is dispensable for the response to H2O2. These observations also suggest functional specialization among cytosolic thioredoxins since another Chlamydomonas isoform (Trxh2) does not compensate for the lack of Trxh1.

Original languageEnglish (US)
Pages (from-to)262-273
Number of pages12
JournalEukaryotic Cell
Volume4
Issue number2
DOIs
StatePublished - Feb 1 2005

Fingerprint

Thioredoxin h
methyl methanesulfonate
alkylation
Chlamydomonas reinhardtii
Alkylation
Methyl Methanesulfonate
DNA damage
DNA Damage
Chlamydomonas
DNA repair
hypersensitivity
Thioredoxins
Bleomycin
DNA Repair
mutants
Hypersensitivity
lesions (plant)
gene deletion
organisms
DNA

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Functional specialization of Chlamydomonas reinhardtii cytosolic thioredoxin h1 in the response to alkylation-induced DNA damage. / Sarkar, Nandita; Lemaire, Stéphane; Wu-Scharf, Danxia; Issakidis-Bourguet, Emmanuelle; Cerutti, Heriberto.

In: Eukaryotic Cell, Vol. 4, No. 2, 01.02.2005, p. 262-273.

Research output: Contribution to journalArticle

Sarkar, Nandita ; Lemaire, Stéphane ; Wu-Scharf, Danxia ; Issakidis-Bourguet, Emmanuelle ; Cerutti, Heriberto. / Functional specialization of Chlamydomonas reinhardtii cytosolic thioredoxin h1 in the response to alkylation-induced DNA damage. In: Eukaryotic Cell. 2005 ; Vol. 4, No. 2. pp. 262-273.
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