Signal transduction triggered by iron to induce the nuclear importation of a Myb3 transcription factor in the parasitic protozoan Trichomonas vaginalis

Hong Ming Hsu, Yu Lee, Pang Hung Hsu, Hsing Wei Liu, Chien Hsin Chu, Ya Wen Chou, Yet Ran Chen, Shu-Hui Chen, Jung Hsiang Tai

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Iron was previously shown to induce rapid nuclear translocation of a Myb3 transcription factor in the protozoan parasite, Trichomonas vaginalis. In the present study, iron was found to induce a transient increase in cellular cAMP, followed by the nuclear influx of Myb3, whereas the latter was also induced by 8-bromo-cyclic AMP. Iron-inducible cAMP production and nuclear influx of Myb3 were inhibited by suramin and SQ22536, respective inhibitors of the Gα subunit of heterotrimeric G proteins and adenylyl cyclases. In contrast, the nuclear influx of Myb3 induced by iron or 8-bromo-cAMP was delayed or inhibited, respectively, by H89, the inhibitor of protein kinase A. Using liquid chromatography-coupled tandem mass spectrometry, Thr156 and Lys143 in Myb3 were found to be phosphorylated and ubiquitinated, respectively. These modifications were induced by iron and inhibited by H89, as shown by immunoprecipitation-coupled Western blotting. Iron-inducible ubiquitination and nuclear influx were aborted in T156A and K143R, but T156D was constitutively ubiquitinated and persistently localized to the nucleus. Myb3 was phosphorylated in vitro by the catalytic subunit of a T. vaginalis protein kinase A, TvPKAc. A transient interaction between TvPKAc and Myb3 and the phosphorylation of both proteins were induced in the parasite shortly after iron or 8-bromo-cAMP treatment. Together, these observations suggest that iron may induce production of cAMP and activation of TvPKAc, which then induces the phosphorylation of Myb3 and subsequent ubiquitination for accelerated nuclear influx. It is conceivable that iron probably exerts a much broader impact on the physiology of the parasite than previously thought to encounter environmental changes.

Original languageEnglish
Pages (from-to)29334-29349
Number of pages16
JournalJournal of Biological Chemistry
Volume289
Issue number42
DOIs
Publication statusPublished - 2014 Oct 17

Fingerprint

Trichomonas vaginalis
Signal transduction
Signal Transduction
Transcription Factors
Iron
8-Bromo Cyclic Adenosine Monophosphate
Parasites
Phosphorylation
Ubiquitination
Cyclic AMP-Dependent Protein Kinases
Heterotrimeric GTP-Binding Proteins
Suramin
Liquid chromatography
Physiology
Tandem Mass Spectrometry
Immunoprecipitation
Adenylyl Cyclases
Liquid Chromatography
Mass spectrometry
Catalytic Domain

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hsu, Hong Ming ; Lee, Yu ; Hsu, Pang Hung ; Liu, Hsing Wei ; Chu, Chien Hsin ; Chou, Ya Wen ; Chen, Yet Ran ; Chen, Shu-Hui ; Tai, Jung Hsiang. / Signal transduction triggered by iron to induce the nuclear importation of a Myb3 transcription factor in the parasitic protozoan Trichomonas vaginalis. In: Journal of Biological Chemistry. 2014 ; Vol. 289, No. 42. pp. 29334-29349.
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abstract = "Iron was previously shown to induce rapid nuclear translocation of a Myb3 transcription factor in the protozoan parasite, Trichomonas vaginalis. In the present study, iron was found to induce a transient increase in cellular cAMP, followed by the nuclear influx of Myb3, whereas the latter was also induced by 8-bromo-cyclic AMP. Iron-inducible cAMP production and nuclear influx of Myb3 were inhibited by suramin and SQ22536, respective inhibitors of the Gα subunit of heterotrimeric G proteins and adenylyl cyclases. In contrast, the nuclear influx of Myb3 induced by iron or 8-bromo-cAMP was delayed or inhibited, respectively, by H89, the inhibitor of protein kinase A. Using liquid chromatography-coupled tandem mass spectrometry, Thr156 and Lys143 in Myb3 were found to be phosphorylated and ubiquitinated, respectively. These modifications were induced by iron and inhibited by H89, as shown by immunoprecipitation-coupled Western blotting. Iron-inducible ubiquitination and nuclear influx were aborted in T156A and K143R, but T156D was constitutively ubiquitinated and persistently localized to the nucleus. Myb3 was phosphorylated in vitro by the catalytic subunit of a T. vaginalis protein kinase A, TvPKAc. A transient interaction between TvPKAc and Myb3 and the phosphorylation of both proteins were induced in the parasite shortly after iron or 8-bromo-cAMP treatment. Together, these observations suggest that iron may induce production of cAMP and activation of TvPKAc, which then induces the phosphorylation of Myb3 and subsequent ubiquitination for accelerated nuclear influx. It is conceivable that iron probably exerts a much broader impact on the physiology of the parasite than previously thought to encounter environmental changes.",
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Signal transduction triggered by iron to induce the nuclear importation of a Myb3 transcription factor in the parasitic protozoan Trichomonas vaginalis. / Hsu, Hong Ming; Lee, Yu; Hsu, Pang Hung; Liu, Hsing Wei; Chu, Chien Hsin; Chou, Ya Wen; Chen, Yet Ran; Chen, Shu-Hui; Tai, Jung Hsiang.

In: Journal of Biological Chemistry, Vol. 289, No. 42, 17.10.2014, p. 29334-29349.

Research output: Contribution to journalArticle

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AU - Hsu, Hong Ming

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AU - Chu, Chien Hsin

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AU - Chen, Shu-Hui

AU - Tai, Jung Hsiang

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