Progress 12/08/15 to 09/30/20
Outputs
Target Audience:General Scientific Community - National and International General Public - Local, State, and National Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has led to the training and professional development of fourgraduate students Nina Serratore, Kortany Baker, Debasmita Saha and Smriti Hoda. These students received training in performing, writing and commutating our results as a publication and at conferences. They also developed abilities to critically analyze results. Nina Serratore also graduated with her PhDand obtained a job in industry. In addition, this project also trained fourundergraduate students. Theirwork will contributeto a published manuscript. In addition, they attend laboratory meetings and they also have to write a written report at the end of each semester. They also can obtain summer and academic fellowships to perform paidresearch in the lab and for professional development for research or medical careers. How have the results been disseminated to communities of interest?Our results of this project havebeen published. In addition, our work has also been presented at a scientific conference (Candida and Candidiasis Conference). Our work would have been presented at other scientific conference but due to Covid19 all conferences where cancelled. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
For this project we accomplished a majority if not all of the goals. Goal 1: We determined that Set4 expression is induced by the sterol sensing transcriptional activators, Upc2 and Ecm22 under hypoxic conditions and was published in Genetics in 2018 (Serratore 2018). Goal 2 was also published (Serratore 2018) where we determine that Set4 functions as a corepressor for the Hap1 transcription factor to repress ergosterol biosynthesis genes under anaerobic growth conditions. Goal 3 we determined by RNA-sequencing analysis and chromatin immunoprecipitation that Set4 targets ERG genes under anaerobic conditions and is recruited to these genes by the Hap1 transcription factor. We also demonstrate that Set4 expression is upregulated in an erg3 deletion strain suggesting a precursor sterol, but not ergosterol, regulates Set4 levels. Overall, we have discovered a new sterol-Upc2 signaling pathway mediated by Set4 that governs azole drug efficacy and sterol homeostasis under hypoxic conditions. These observations were published in Genetics (Serratore 2018). Goal 4: We have generated spontaneous suppressors for genomic mutations when Set4 is overexpressed. We will be sequencing these strains for identification of the suppressor mutations. We are also characterizing the role of Set1, Set4 and Hap1 in a human fungal pathogen Candida glabrata. Interesting, they all are necessary for ergosterol homeostasis upon azole drug treatment.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2020
Citation:
Strahl BD and Briggs SD. The SAGA continues: The rise of cis- and trans-histone crosstalk pathways. Biochim Biophys Acta Gene Regul Mech. 2020 Jul 6:194600. doi: 10.1016/j.bbagrm.2020.194600.
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Progress 10/01/18 to 09/30/19
Outputs
Target Audience:General Scientific Community - National and International General Public - Local, State, and National Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has led to the training and professional development of two graduate student Kortany Baker and Smrithi Hoda. These students received training in performing, writing and commicating our results as a publication and at conferences. They also developed abilities to critically anazye results. Nina Serratore also graduated and obtained a job in industry. In addtion, this project also trained three undergraduate student which two willcontributed to a published manuscript to be submitted this semester. How have the results been disseminated to communities of interest?Our results of this project will be sumbitted for publication this semester. In addition, our work will also presented at a scientific conference (Candida and Candidiasis Conference, Montreal, Canada). What do you plan to do during the next reporting period to accomplish the goals?We also plan to published a manuscript this fall "The impact of H3H4 methylation and ergosterol homeostasis on antifungal azole drug resistance in Candida glabrata". In addition, next semester we plan to publish another paper on how epigenetics impact oxidative stress.
Impacts
What was accomplished under these goals?
We accomplished various aspect of goals 1 and 3 of this project. Goal 1: We determined that Set4 expression is induced by the sterol sensing transcriptional activators, Upc2 and Ecm22 under hypoxic conditions. Goal 3: In addtion, we determined by RNA-sequencing analysis that Set4 is required for ERG gene repression. Importantly, we show that Set4 directly targets ergosterol gene promoters, ERG11 and ERG3 under hypoxia and that Set4 recruitment is dependent on the transcriptional repressor, Hap1. Finally, we demonstrate that Set4 expression is upregulated in an erg3 deletion strain suggesting a precursor sterol, but not ergosterol, regulates Set4 levels. Overall, we have discovered a new sterol-Upc2 signaling pathway mediated by Set4 that governs azole drug efficacy and sterol homeostasis under hypoxic conditions. These observations were published in Genetics (2018). Goal 2: We are continuing to determine the biochemical function/methyltranferase function of Set4 and Set1. Goal 4: We will sequence our spontaneous suppressors for genomic mutations when Set4 is overexpressed. We are also characterizing the role of Set1, Set4 and Hap1in a human funal pathogen Candida glabrata.
Publications
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Progress 10/01/17 to 09/30/18
Outputs
Target Audience:General Scientific Community - National and International General Public - Local, State, and National Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has led to the training and professional development of two graduate student Nina Serratore and Kortany Backer. These students received training in performing, writing and commicating our results as a publication and at conferences. They also developed abilities to critically anazye results. Nina Serratore also graduated and obtained a job in industry. In addtion, this project also trained three undergraduate student which also contributed to pur published manuscript. How have the results been disseminated to communities of interest?Our results of this project have published in Genetics, a peer-reviewed science journal. In addition, our work was also presented at a scientific conference (14th ASM Candida and Candidiasis Conference). What do you plan to do during the next reporting period to accomplish the goals?We plan to have our manuscript published "Determining how Set1 interacts with RNA polymerase." We also plan to published an additional manuscript "The impact of H3H4 methylation and ergosterolhomeostasis on antifungal azole drug resistance in Candida glabrata"
Impacts
What was accomplished under these goals?
We accomplished various aspect of goals 1 and 3 of this project. Goal 1: We determined that Set4 expression is induced by the sterol sensing transcriptional activators, Upc2 and Ecm22 under hypoxic conditions. Goal 3: In addtion, we determined by RNA-sequencing analysis that Set4 is required for ERG gene repression. Importantly, we show that Set4 directly targets ergosterol gene promoters, ERG11 and ERG3 under hypoxia and that Set4 recruitment is dependent on the transcriptional repressor, Hap1. Finally, we demonstrate that Set4 expression is upregulated in an erg3 deletion strain suggesting a precursor sterol, but not ergosterol, regulates Set4 levels. Overall, we have discovered a new sterol-Upc2 signaling pathway mediated by Set4 that governs azole drug efficacy and sterol homeostasis under hypoxic conditions. These observations were published in Genetics (2018). Goal 2: We are still continuing to determine the biochemical function/methyltranferase function of Set4. Goal 4: Currently, we have identified spontaneous suppress when Set4 is overexpressed. These suppressors are being evaulated biochemically and for genomic mutations.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Serratore, N.D., Baker, K.M., Macadlo, L.A., Gress, A.R., Powers, B.L., Atallah, N., Westerhouse K.M., Hall, M.C., Weake, V.M., Briggs, S.D. A Novel Sterol-Signaling Pathway Governs Azole Antifungal Drug Resistance and Hypoxic Gene Repression in Saccharomyces cerevisiae. Genetics. 208: 1037-1055, 2018. doi: 10.1534/genetics.117.300554.
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience:General Scientific Community - Nationally and Internationally General Public - Local, State, and National Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has led to the training and professional development of two graduate students Nina Serratore and Kortany Baker. These students received training in performing, writing and commicating our results as amanuscript under and at conferences. They also developed abilities to critically anazye results. In addtion, this project also trained three undergraduate students which also contributed to authorship on a manuscript. How have the results been disseminated to communities of interest?Our results of this project will be published in peer-reviewed science journals. In addition, our work will also be presented at scientific conferences. What do you plan to do during the next reporting period to accomplish the goals?We plan to have our manuscript published"Sterol facilitated induction of Set4 governs antifungal drug efficacy and mediates epigenetic changes under hypoxic conditions in Saccharomyces cerevisiae".We also plan to published an additional manuscript "Determining how Set1 interacts with RNA polymerase."
Impacts
What was accomplished under these goals?
We accomplished various aspect of goals 1 and 3of this project. Goal 1: Wedetermined that Set4 expression is induced by the sterol sensing transcriptional activators, Upc2 and Ecm22. Goal 3: In addtion, we determined by RNA-sequencing analysis that Set4 is required for ERG gene repression. Importantly, we show that Set4 directly targets ergosterol gene promoters, ERG11 and ERG3 under hypoxia and that Set4 recruitment is dependent on the transcriptional repressor, Hap1. Finally, we demonstrate that Set4 expression is upregulated in an erg3? strain suggesting a precursor sterol, but not ergosterol, regulates Set4 levels. Overall, we have discovered a new sterol-Upc2 signaling pathway mediated by Set4 that governs azole drug efficacy and sterol homeostasis under hypoxic conditions.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Zhang, Y., Serratore, N.D., and Briggs, S.D. N-ICE plasmids for generating N-terminal 3?�?FLAG tagged genes that allow inducible, constitutive or endogenous expression in Saccharomyces cerevisiae. Yeast. 34: 223-235, 2017.
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Progress 12/08/15 to 09/30/16
Outputs
Target Audience:General Scientific Community -- Nationally and Internationally General Public - Local, State, and Nationally Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has led to the training and professional development of two graduate students Nina Serratore andYueping Zhang. These students received training in performing, writing and commicating our results as a publication and at conferences. They also developed abilities to critically anazye results. In addtion, this project also trained two undergraduate students. How have the results been disseminated to communities of interest?Our results of this project will be published in peer-reviewed sciencejournals. In addition,our work has also been presented at scientific conferences. What do you plan to do during the next reporting period to accomplish the goals?We plan to have our manuscript published "N-ICE plasmids for generating N-terminal 3×FLAG tagged genes that allow Inducible, Constitutive, or Endogenous expression in Saccharomyces cerevisiae." We also plan to published two additional manuscripts: Manuscript 1) Determining how Set1 interacts with RNA polymerase and 2) Determining the biological and biochemical function of Set4.
Impacts
What was accomplished under these goals?
One of the goals that was accomplished was the development of a newplasmids and methodto tag proteins in yeast. We have constructed the N-ICE plasmid system for PCR-based N-terminal 3×FLAG epitope tagging of non-essential and essential genes under the control an Inducible promoter (GAL1), Constitutive promoters (CYC1 or PYK1), or the Endogenous promoter. Our N-ICE plasmid system also provides the option of three different selection markers, including KanMX, HphMX, and NatMX cassettes. In addition, the integrated selection marker and the heterologous promoter can be excised with a subsequent step using Cre recombinase, leaving the target gene expressed from the endogenous promoter. This system can also be applied to tagging essential genes expressed by their endogenous promoters. We have validated the N-ICE system by N-terminal tagging two non-essential genes (SET1 and SET2) and two essential genes (ERG11 and PKC1). Overall, the N-ICE plasmid system will provide valuable new tools to enhance the yeast epitope tagging toolbox.These new "N-ICE" plasmids were constructed so that non-essential and essential genes can be N-terminally 3×FLAG tagged and expressed from an Inducible promoter (GAL1), Constitutive promoters (CYC1 or PYK1), or the Endogenous promoter. The second goal that was accomplished was determining how Set1 interacted with RNA Polymerase II.Currently, this work is being writteninto a manuscript which we hope to have published next year. The other goals which are still in progress are determining how Set4 is regulated, Defining the biochemical function of Set4, Identification of Set4 target genes, and identification of suppressors for azole resistant strains. These remaining goals are still ongoing in the laboratory.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2017
Citation:
Zhang, Y., Serratore, N.D., Briggs, S.D. N-ICE plasmids for generating N-terminal 3�FLAG tagged genes that allow Inducible, Constitutive, or Endogenous expression in Saccharomyces cerevisiae. Submitted and underreview 2016
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