Yeast adenylate kinase is transcribed constitutively from a promoter in the short intergenic region to the histone H2A‐1 gene

Oechsner, Ulrich; Magdolen, Viktor; Zoglowek, Cornelia; Häcker, Udo; Bandlow, Wolfhard

doi:10.1016/0014-5793(88)81013-5

Cited by 18 publications

(5 citation statements)

References 33 publications

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“…2) analysis contains both HTAJ and AKY2IPRTI DNA sequences. The latter is a contiguous gene which codes for an mRNA that is not cell cycle regulated and serves as an internal control (32,35). RNA taken from unarrested wildtype cells (HPC+) produces an HTAI band which is considerably darker than the PRTJ mRNA band (lane 1).…”

Section: Resultsmentioning

confidence: 99%

Identification of a new set of cell cycle-regulatory genes that regulate S-phase transcription of histone genes in Saccharomyces cerevisiae.

Xu¹,

Kim²,

Schuster³

et al. 1992

Mol. Cell. Biol.

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Histone mRNA synthesis is tightly regulated to S phase of the yeast Saccharomyces cerevisiae cell cycle as a result of transcriptional and posttranscriptional controls. Moreover, histone gene transcription decreases rapidly if DNA replication is inhibited by hydroxyurea or if cells are arrested in G1 by the mating pheromone a-factor. To identify the transcriptional controls responsible for cycle-specific histone mRNA synthesis, we have developed a selection for mutations which disrupt this process. Using this approach, we have isolated five mutants (hpcl, hpc2, hpc3, hpc4, and Histone mRNAs are synthesized and accumulated preferentially in S phase of the eukaryotic cell cycle, partly as a result of transcriptional controls which cause the rate of synthesis of histone mRNAs to increase during S phase. Also, posttranscriptional regulation results in the preferential degradation of histone mRNAs outside of S (reviewed in reference 17). These two mechanisms of regulation function to decrease mammalian histone mRNA levels after DNA synthesis is blocked in the presence of hydroxyurea (HU) (17). In this report, we focus on cell cycle-specific, transcriptional control of histone mRNA synthesis.The human histone H4 gene is transcribed more efficiently (3-to 10-fold) in S-phase extracts than in extracts taken from cells at other stages of the cell cycle (18). Mammalian upstream promoter elements responsible for this regulation have been identified (1,6,16,28,42). Moreover, fusion of the 5' end of the mouse histone H3 gene to the bacterial neomycin resistance (neo) gene causes the bacterial mRNA to be synthesized in a cell cycle-specific manner (3). cisacting regulatory sequences and their trans-acting protein factors are likely responsible for regulating histone mRNA synthesis (7, 10, 11, 14, 50; reviewed in reference 33); however, the molecular details of the regulatory mechanism by which this occurs are still under investigation.The yeast Saccharomyces cerevisiae provides a system which allows the identification genetically of trans-acting factors that regulate cycle-specific histone mRNA synthesis. Yeast

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Section: Resultsmentioning

confidence: 99%

Identification of a new set of cell cycle-regulatory genes that regulate S-phase transcription of histone genes in Saccharomyces cerevisiae.

Xu¹,

Kim²,

Schuster³

et al. 1992

Mol. Cell. Biol.

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“…SPK1 mRNA, initially low, was maximal at 10 or 20 min in the first cycle after release and appeared in the interval between appearance of nonbudded cells and smallbudded cells in each cycle. LEU2 and protein 1 (43) mRNAs, used as internal controls, were reasonably constant throughout the experiment. SPKI mRNA accumulation correlated with that of RFA2, an MCB-regulated gene (6).…”

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confidence: 99%

SPK1 is an essential S-phase-specific gene of Saccharomyces cerevisiae that encodes a nuclear serine/threonine/tyrosine kinase.

et al. 1993

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SPKI was originally discovered in an immunoscreen for tyrosine-protein kinases in Saccharomyces cerevisiae. MATERIALS AND METHODS Plasmids. pSK9-15 and pSK15-9 are Bluescript plasmids (Stratagene Cloning Systems) carrying a 3.6-kbp EcoRI fragment including SPK1 coding and upstream sequences (52); pNB187 was obtained from Peter Novick (Yale University School of Medicine); clnl::URA3 and pH2AB1 were obtained from Fred Cross (Rockefeller University); pRS303, pRS305, pRS315, and pRS316 (51) were obtained from Aaron Mitchell (College of Physicians and Surgeons, Columbia University); and pJM229 (6) was obtained from Steve Brill (Rutgers University).pNB187-SPKI. The 2,764-bp HindIll fragment delineated by the single HindIII site upstream from the SPK1 coding sequences and the Bluescript polylinker (downstream from the SPK1 insert) in pSK9-15 was cloned into the BamHI site of pNB187 by using BamHI-HindIII adaptors. In the resulting plasmid, pNB187-SPK1, SPK1 coding sequences follow the GALl promoter and are flanked by functional BamHI, but not HindIII, cleavage sites. pNB187-spklK227A. pSK9-15 was used as a template for four-primer polymerase chain reaction mutagenesis to mutate nucleotides 712 to 717 (original numbering system [52]) from AAG (Lys) ATT (Ile) to GCA (Ala) ATC (Ile). First, two reactions were run in parallel to produce overlapping DNA fragments with one mutation as follows: reaction A, wild-type forward primer (5') CCCAGCCTGTGACTATCA 'Tl r7AGG (3') and mutagenic reverse primer (5') GCGTTTA ClGTATGATTGCCACC (3'); reaction B, mutagenic forward 5829

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“…Expression of these histone genes has been shown to occur periodically in the cell cycle in a start-dependent manner (13). Located adjacent to the cloned TRTJ locus on the same DNA insert is the protein 1 (adenylate kinase) (19,29) gene that, unlike the neighboring histone genes, shows constant expression throughout the cell cycle (13). Our initial intention was to use this probe to indicate both increased biosynthetic activity, evidenced by protein 1 transcript levels, and cell cycle activity, evidenced by histone transcript levels.…”

Section: Resultsmentioning

confidence: 99%

Induction of yeast histone genes by stimulation of stationary-phase cells.

Drebot¹,

Veinot-Drebot²,

Singer³

et al. 1990

Mol. Cell. Biol.

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In the cell cycle of the budding yeast Saccharomyces cerevisiae, expression of the histone genes H2A and H2B of the TRTI and TRT2 loci is regulated by the performance of "start," the step that also regulates the cell cycle. Here we show that histone production is also subject to an additional form of regulation that is unrelated to the mitotic cell cycle. Expression of histone genes, as assessed by Northern (RNA) analysis, was shown to increase promptly after the stimulation, brought about by fresh medium, that activates stationary-phase cells to reenter the mitotic cell cycle. The use of a yeast mutant that is conditionally blocked in the resumption of proliferation at a step that is not part of the mitotic cell cycle (M. A. Drebot, G. C. Johnston, and R. A. Singer, Proc. Natl. Acad. Sci. 84:7948, 1987) (11,13). During the mitotic cell cycle, the expression of these histone genes occurs periodically, with significant accumulation of histone mRNA after performance of the cell cycle regulatory step termed "start" (9) and with maximum levels achieved in the S phase (13). Cells treated with yeast mating pheromone to block performance of start do not contain significant levels of H2A and H2B mRNA (13).Here we show by analysis of transcript levels in both wild-type and mutant cells (3, MATERIALS AND METHODSStrains and plasmids. The wild-type strain S. cerevisiae GR2 (MATa ural his6) has been described previously (17). Strain MD-G3G02 (MATa gcsl-J sedl-J ade6 ural his6) is a yeast mutant that is conditionally defective only for resumption of proliferation from the stationary phase (3, 4). Plasmids YIp5-TRT1 (32) and

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Yeast adenylate kinase is transcribed constitutively from a promoter in the short intergenic region to the histone H2A‐1 gene

Cited by 18 publications

References 33 publications

Identification of a new set of cell cycle-regulatory genes that regulate S-phase transcription of histone genes in Saccharomyces cerevisiae.

Identification of a new set of cell cycle-regulatory genes that regulate S-phase transcription of histone genes in Saccharomyces cerevisiae.

SPK1 is an essential S-phase-specific gene of Saccharomyces cerevisiae that encodes a nuclear serine/threonine/tyrosine kinase.

Induction of yeast histone genes by stimulation of stationary-phase cells.

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