
Amanda GuiseBiogen · Translational Neuropathology
Amanda Guise
PhD
About
33
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September 2009 - June 2015
Publications
Publications (33)
To elucidate the role of Tau isoforms and post-translational modification (PTM) stoichiometry in Alzheimer’s disease (AD), we generated a high-resolution quantitative proteomics map of 95 PTMs on multiple isoforms of Tau isolated from postmortem human tissue from 49 AD and 42 control subjects. Although Tau PTM maps reveal heterogeneity across subje...
Alternative translation initiation and stop codon readthrough in a few well-studied cases have been shown to allow the same transcript to generate multiple protein variants. Because the brain shows a particularly abundant use of alternative splicing, we sought to study alternative translation in CNS cells. We show that alternative translation is wi...
Translation canonically begins at a single AUG and terminates at the stop codon, generating one protein species per transcript. However, some transcripts may use alternative initiation sites or sustain translation past their stop codon, generating multiple protein isoforms. Through other mechanisms such as alternative splicing, both neurons and gli...
Replicative DNA helicases expose the two strands of the double helix to the replication apparatus, but accessory helicases are often needed to help forks move past naturally occurring hard-to-replicate sites, such as tightly bound proteins, RNA/DNA hybrids, and DNA secondary structures. Although the Schizosaccharomyces pombe 5'-to-3' DNA helicase P...
Fork pausing and DNA damage are increased at G4 motifs in the absence of Pfh1.
(A) Scatter plot comparing Cdc20 peak strength (-10*log10(p-value)) at G4 motifs in WT and Pfh1-depleted cells. Each point represents a genomic region with a Cdc20 occupancy peak in at least one context. If a peak was not present in a context, it is plotted at 0 on the c...
Interaction of Pfh1-GFP with DNA replication proteins Pol2 and the Mcm helicase complex observed in asynchronous cells.
Immunoaffinity-purification of Pfh1-GFP from asynchronous cells. Proteins were resolved by SDS-PAGE and visualized by Coomassie stain. Peptides of identified proteins were confirmed by MS with nanoLC LTQ Orbitrap CID analyses. Pfh...
Immunoaffinity-purification of Pfh1-GFP is not affected by DNaseI treatment of the cell lysate.
(A) Proteins were resolved by SDS-PAGE and visualized by Coomassie stain. (B) Ethidium bromide stained agarose gel of precipitated DNA from an aliquot of the cell lysate before and after DNAseI treatment (lanes 1 and 2) and with the addition of plasmid D...
(A) Fraction of Pfh1-bound features of a given type that overlap Pfh1-bound instances of each other tested feature type. (B) Fraction of Pfh1-sensitive (in terms of Cdc20) features of a given type that overlap Pfh1-bound instances of each other tested feature type. In each panel, the fraction is given in terms of the y-axis feature. For example, th...
Analysis of Pfh1-binding at sites that are sensitive to the depletion of Pfh1.
(XLSX)
Pfh1, Cdc20, and ?-H2A signals surrounding all 5S rRNA genes.
Details are as in Fig 3.
(PDF)
Proteins Interacting with Pfh1 in either S or G2 Phase Identified by Immunoaffinity Purification Mass Spectrometry.
(XLSX)
S. pombe strains used in this study.
(DOCX)
Pfh1 is bound to all DNA regions tested.
Samples from asynchronous cells either expressing Pfh1-13Myc or an untagged control were chromatin immunoprecipitated using an anti-Myc antibody. Data for Pfh1-13Myc were also shown in Fig 1. The immunoprecipitated DNA was quantified using quantitative PCR with primers specific for hsp90+, tdh1+, adh1+, hta1...
Pfh1, Cdc20, and ?-H2A signals surrounding all tRNA genes.
Details are as in Fig 3.
(PDF)
Genome-wide Peak Locations for Pfh1, Cdc20 in WT, Cdc20 in Pfh1-depleted cells, ?-H2A in WT, and ?-H2A in Pfh1-depleted Cells.
(XLSX)
Cdc20 and ?-H2A Peaks Are Stronger in Pfh1-depleted Cells than WT Cells.
(XLSX)
Genomic Features Associated with ?-H2A Occupied Regions in WT and Pfh1-depleted Cells.
(XLSX)
Genomic Features Associated with shared and unique Cdc20 and ?-H2A Occupied Regions.
(XLSX)
Oligonucleotides used for qPCR experiments.
(DOCX)
Genomic Features Associated with Cdc20 Occupied Regions in WT and Pfh1-depleted Cells.
(XLSX)
As a member of the class IIa family of histone deacetylases, the histone deacetylase 5 (HDAC5) is known to undergo nuclear–cytoplasmic shuttling and to be a critical transcriptional regulator. Its misregulation has been linked to prominent human diseases, including cardiac diseases and tumorigenesis. In this chapter, we describe several experimenta...
In biological systems, proteins catalyze the fundamental reactions that underlie all cellular functions, including metabolic processes and cell survival and death pathways. These biochemical reactions are rarely accomplished alone. Rather, they involve a concerted effect from many proteins that may operate in a directed signaling pathway and/or may...
Deleted in breast cancer 1 (DBC1) has emerged as an important regulator of multiple cellular processes, ranging from gene expression to cell cycle progression. DBC1 has been linked to tumorigenesis both as an inhibitor of histone deacetylases (HDAC), HDAC3 and sirtuin 1 (SIRT1), and as a transcriptional cofactor for nuclear hormone receptors. Howev...
Almost 400 genes affect yeast telomere length, including Est1, which is critical for recruitment and activation of telomerase. Here we use mass spectrometry to identify novel telomerase regulators by their co-purification with the telomerase holoenzyme. In addition to all known subunits, over 100 proteins are telomerase associated, including all th...
Class IIa histone deacetylases (HDACs4, -5, -7, and -9) modulate the physiology of the human cardiovascular, musculoskeletal, nervous and immune systems. The regulatory capacity of this family of enzymes stems from their ability to shuttle between nuclear and cytoplasmic compartments in response to signal-driven post-translational modification. Her...
Class IIa histone deacetylases (HDACs) are critical transcriptional regulators, shuttling between nuclear and cytoplasmic cellular compartments. Within the nucleus, these HDACs repress transcription as components of multi-protein complexes, such as the nuclear co-repressor (NCoR) and beclin-6 co-repressor (BCoR) complexes. Cytoplasmic relocalizatio...
Emerging evidence highlights a critical role for protein acetylation during herpesvirus infection. As prominent modulators of protein acetylation, histone deacetylases (HDACs) are essential transcriptional and epigenetic regulators. Not surprisingly, viruses have evolved a wide array of mechanisms to subvert HDAC functions. Here, we review the mech...
Histone deacetylases (HDACs) are a diverse family of essential transcriptional regulatory enzymes, that function through the spatial and temporal recruitment of protein complexes. As the composition and regulation of HDAC complexes are only partially characterized, we built the first global protein interaction network for all 11 human HDACs in T ce...
Class IIa histone deacetylases (HDACs 4/5/7/9) are transcriptional regulators with critical roles in cardiac disease, cancer, and viral infection. HDAC inhibitors are promising anti-cancer agents, and while they are known to disrupt mitotic progression, the underlying mechanisms of mitotic regulation by HDACs are not fully understood. Here we provi...
Histone deacetylase 5 (HDAC5), a class IIa deacetylase, is a prominent regulator of cellular and epigenetic processes that underlie the progression of human disease, ranging from cardiac hypertrophy to cancer. Although it is established that phosphorylation mediates 14-3-3 protein binding and provides the essential link between HDAC5 nucleo-cytopla...