Gln3p Nuclear Localization and Interaction with Ure2p in Saccharomyces cerevisiae

Ajit A. Kulkarni, Ashraf T. Abul-Hamd, Rajendra Rai, Hassan El Berry, Terrance Cooper

Research output: Contribution to journalArticle

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Abstract

Gln3p is one of two well characterized GATA family transcriptional activation factors whose function is regulated by the nitrogen supply of the cell. When nitrogen is limiting, Gln3p and Gat1p are concentrated in the nucleus where they bind GATA sequences upstream of nitrogen catabolite repression (NCR)-sensitive genes and activate their transcription. Conversely, in excess nitrogen, these GATA sequences are unoccupied by Gln3p and Gat1p because these transcription activators are excluded from the nucleus. Ure2p binds to Gln3p and Gat1p and is required for NCR-sensitive transcription to be repressed and for nuclear exclusion of these transcription factors. Here we show the following. (i) Gln3p residues 344-365 are required for nuclear localization. (ii) Replacing Ser-344, Ser-347, and Ser-355 with alanines has minimal effects on GFP-Gln3p localization. However, replacing Gln3p Ser-344, Ser-347, and Ser-355 with aspartates results in significant loss of its ability to be concentrated in the nucleus. (iii) N and C termini of the Gln3p region required for it to complex with Ure2p and be excluded from the nucleus are between residues 1-103 and 301-365, respectively. (iv) N and C termini of the Ure2p region required for it to interact with Gln3p are situated between residues 101-151 and 330-346, respectively. (v) Loss of Ure2p residues participating in either dimer or prion formation diminishes its ability to carry out NCR-sensitive regulation of Gln3p activity.

Original languageEnglish (US)
Pages (from-to)32136-32144
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number34
DOIs
StatePublished - Aug 24 2001

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Yeast
Saccharomyces cerevisiae
Nitrogen
Catabolite Repression
Transcription
Prions
Aspartic Acid
Alanine
Dimers
Transcriptional Activation
Transcription Factors
Genes
Chemical activation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Gln3p Nuclear Localization and Interaction with Ure2p in Saccharomyces cerevisiae. / Kulkarni, Ajit A.; Abul-Hamd, Ashraf T.; Rai, Rajendra; El Berry, Hassan; Cooper, Terrance.

In: Journal of Biological Chemistry, Vol. 276, No. 34, 24.08.2001, p. 32136-32144.

Research output: Contribution to journalArticle

Kulkarni, Ajit A. ; Abul-Hamd, Ashraf T. ; Rai, Rajendra ; El Berry, Hassan ; Cooper, Terrance. / Gln3p Nuclear Localization and Interaction with Ure2p in Saccharomyces cerevisiae. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 34. pp. 32136-32144.
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