Donna E. Crone
University of California, Riverside
Network
Latest external collaboration on country level. Dive into details by clicking on the dots.
Publication
Featured researches published by Donna E. Crone.
Cancer Research | 2007
Satish Sankaran; Donna E. Crone; Robert E. Palazzo; Jeffrey D. Parvin
Breast cancer-associated gene 1 (BRCA1) regulates the duplication and the function of centrosomes in breast cells. We have previously shown that BRCA1 ubiquitin ligase activity directly inhibits centrosome-dependent microtubule nucleation. However, there is a paradox because centrosome microtubule nucleation potential is highest during mitosis, a phase when BRCA1 is most abundant at the centrosome. In this study, we resolve this conundrum by testing whether centrosomes from cells in M phase are regulated differently by BRCA1 when compared with other phases of the cell cycle. We observed that BRCA1-dependent inhibition of centrosome microtubule nucleation was high in S phase but was significantly lower during M phase. The cell cycle-specific effects of BRCA1 on centrosome-dependent microtubule nucleation were detected in living cells and in cell-free experiments using centrosomes purified from cells at specific stages of the cell cycle. We show that Aurora-A kinase modulates the BRCA1 inhibition of centrosome function by decreasing the E3 ubiquitin ligase activity of BRCA1. In addition, dephosphorylation of BRCA1 by protein phosphatase 1 alpha enhances the E3 ubiquitin ligase activity of BRCA1. These observations reveal that the inhibition of centrosome microtubule nucleation potential by the BRCA1 E3 ubiquitin ligase is controlled by Aurora-A kinase and protein phosphatase 1 alpha-mediated phosphoregulation through the different phases of the cell cycle.
Cancer Biology & Therapy | 2007
Satish Sankaran; Donna E. Crone; Robert E. Palazzo; Jeffrey D. Parvin
Centrosomes are the cellular organelles that nucleate microtubules (MTs) via the activity of gamma-tubulin ring complex(s) (γ-TuRC) bound to the pericentriolar material of the centrosomes. BRCA1, the breast and ovarian cancer specific tumor suppressor, inhibits centrosomal MT nucleation via its ubiquitin ligase activity, and one of the known BRCA1 substrates is the key γ-TuRC component, γ-tubulin. We analyzed the mechanism by which BRCA1 regulates centrosome function using an in vitro reconstitution assay, which includes separately staged steps. Our results are most consistent with a model by which the BRCA1 ubiquitin ligase modifies both γ-tubulin plus a second centrosomal protein that controls localization of γ-TuRC to the centrosome. We suggest that this second protein is an adapter protein or protein complex that docks γ-TuRC to the centrosome. By controlling γ-TuRC localization, BRCA1 appropriately inhibits centrosome function, and loss of BRCA1 would result in centrosome hyperactivity, supernumerary centrosomes and, possibly, aneuploidy.
Molecular and Cellular Endocrinology | 1993
John B. Tillman; Donna E. Crone; Hyoung-soon Kim; Carl N. Sprung; Stephen R. Spindler
We report that the activity of the firefly luciferase (LUC) reporter gene is down-regulated by T3 and T3 receptor (TR) in the CV1 mammalian cell line, which is widely used for studies of TR action. Repression was highly reproducible, T3 and TR dependent, promoter independent, and observed regardless of whether an internal control for transfection efficiency was used. Cotransfections with normal and mutant TRs indicate that the negative T3 response is mediated by sequences within the LUC gene coding region, and is not due to the interaction of TR with a limiting transcription factor. Negative regulation of the LUC reporter was overcome by a strong, cis-linked T3 response element (TRE), but continued in the presence of a TRE of moderate strength. The results described here demonstrate that conclusions drawn from studies of TRE structure and activity performed using the LUC reporter in CV1 cells should be interpreted with caution.
Biochemistry | 2015
Yao-ming Huang; Shounak Banerjee; Donna E. Crone; Christian D. Schenkelberg; Derek J. Pitman; Patrick M. Buck; Christopher Bystroff
Leave-one-out green fluorescent protein (LOOn-GFP) is a circularly permuted and truncated GFP lacking the nth β-strand element. LOO7-GFP derived from the wild-type sequence (LOO7-WT) folds and reconstitutes fluorescence upon addition of β-strand 7 (S7) as an exogenous peptide. Computational protein design may be used to modify the sequence of LOO7-GFP to fit a different peptide sequence, while retaining the reconstitution activity. Here we present a computationally designed leave-one-out GFP in which wild-type strand 7 has been replaced by a 12-residue peptide (HA) from the H5 antigenic region of the Thailand strain of H5N1 influenza virus hemagglutinin. The DEEdesign software was used to generate a sequence library with mutations at 13 positions around the peptide, coding for approximately 3 × 10(5) sequence combinations. The library was coexpressed with the HA peptide in E. coli and colonies were screened for in vivo fluorescence. Glowing colonies were sequenced, and one (LOO7-HA4) with 7 mutations was purified and characterized. LOO7-HA4 folds, fluoresces in vivo and in vitro, and binds HA. However, binding results in a decrease in fluorescence instead of the expected increase, caused by the peptide-induced dissociation of a novel, glowing oligomeric complex instead of the reconstitution of the native structure. Efforts to improve binding and recover reconstitution using in vitro evolution produced colonies that glowed brighter and matured faster. Two of these were characterized. One lost all affinity for the HA peptide but glowed more brightly in the unbound oligomeric state. The other increased in affinity to the HA peptide but still did not reconstitute the fully folded state. Despite failing to fold completely, peptide binding by computational design was observed and was improved by directed evolution. The ratio of HA to S7 binding increased from 0.0 for the wild-type sequence (no binding) to 0.01 after computational design (weak binding) and to 0.48 (comparable binding) after in vitro evolution. The novel oligomeric state is composed of an open barrel.
Biochemistry | 2017
Shounak Banerjee; Christian D. Schenkelberg; Thomas B. Jordan; Julia M. Reimertz; Emily E. Crone; Donna E. Crone; Christopher Bystroff
The autocatalytic maturation of the chromophore in green fluorescent protein (GFP) was thought to require the precise positioning of the side chains surrounding it in the core of the protein, many of which are strongly conserved among homologous fluorescent proteins. In this study, we screened for green fluorescence in an exhaustive set of point mutations of seven residues that make up the chromophore microenvironment, excluding R96 and E222 because mutations at these positions have been previously characterized. Contrary to expectations, nearly all amino acids were tolerated at all seven positions. Only four point mutations knocked out fluorescence entirely. However, chromophore maturation was found to be slower and/or fluorescence reduced in several cases. Selected combinations of mutations showed nonadditive effects, including cooperativity and rescue. The results provide guidelines for the computational engineering of GFPs.
Journal of Biological Chemistry | 1990
Donna E. Crone; Hyoung-Soon Kim; Stephen R. Spindler
Molecular Endocrinology | 1992
Hyoung-soon Kim; Donna E. Crone; C. N. Sprung; John B. Tillman; W. R. Force; Mark D. Crew; P. L. Mote; Stephen R. Spindler
Plant Cell and Environment | 2001
Donna E. Crone; J. Rueda; K. L. Martin; D. A. Hamilton; Joseph P. Mascarenhas
Plant Molecular Biology | 1998
Scott A. Campbell; Donna E. Crone; Toni L. Ceccardi; Timothy J. Close
Protein Science | 2015
Derek J. Pitman; Shounak Banerjee; Stephen Macari; Christopher A. Castaldi; Donna E. Crone; Christopher Bystroff