TY - JOUR
T1 - Expression of the purine biosynthetic enzyme phosphoribosyl formylglycinamidine synthase in neurons
AU - Mangold, Colleen A.
AU - Yao, Pamela J.
AU - Du, Mei
AU - Freeman, Willard M.
AU - Benkovic, Stephen J.
AU - Szpara, Moriah L.
N1 - Funding Information:
We thank the faculty and staff members of the Huck Microscopy Facility at Penn State University for their support and guidance during all the imaging experiments. We also thank Dr. Anthony Pedley, Dr. Vidhi Pareek, and Dr. James Lata of the Benkovic lab for their valuable insights in experimental planning and data interpretation, as well as members of the Szpara lab. C.A.M. is a post-doctoral fellow of the American Heart Association. This work was supported by the startup funds (M.L.S.) from the Pennsylvania State University, the National Institutes of Health (RO1 GM024129, S.J.B.), the American Heart Association (16POST29920001, C.A.M.), a J. Lloyd Huck Biotechnology mini-grant (C.A.M.) from the Pennsylvania State University, and the Intramural Research Programs of the National Institute of Health/National Institute on Aging (P.Y.). The Sv2 monoclonal antibody used in this study was developed by Dr. Kathleen M. Buckley at Harvard Medical School and was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. The authors declare no conflicts of interest. All experiments were conducted in compliance with the ARRIVE guidelines.
Funding Information:
We thank the faculty and staff members of the Huck Microscopy Facility at Penn State University for their support and guidance during all the imaging experiments. We also thank Dr. Anthony Pedley, Dr. Vidhi Pareek, and Dr. James Lata of the Benkovic lab for their valuable insights in experimental planning and data interpretation, as well as members of the Szpara lab. C.A.M. is a post-doctoral fellow of the American Heart Association. This work was supported by the startup funds (M.L.S.) from the Pennsylvania State University, the National Institutes of Health (RO1 GM024129, S.J.B.), the American Heart Association (16POST29920001, C.A.M.), a J. Lloyd Huck Biotechnology mini-grant (C.A.M.) from the Pennsylvania State University, and the Intramural Research Programs of the National Institute of Health/National Institute on Aging (P.Y.). The Sv2 monoclonal antibody used in this study was developed by Dr. Kathleen M. Buckley at Harvard Medical School and was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. The authors declare no conflicts of interest.
Publisher Copyright:
© 2018 International Society for Neurochemistry
PY - 2018/3
Y1 - 2018/3
N2 - Purines are metabolic building blocks essential for all living organisms on earth. De novo purine biosynthesis occurs in the brain and appears to play important roles in neural development. Phosphoribosyl formylglycinamidine synthase (FGAMS, also known as PFAS or FGARAT), a core enzyme involved in the de novo synthesis of purines, may play alternative roles in viral pathogenesis. To date, no thorough investigation of the endogenous expression and localization of de novo purine biosynthetic enzymes has been conducted in human neurons or in virally infected cells. In this study, we characterized expression of FGAMS using multiple neuronal models. In differentiated human SH-SY5Y neuroblastoma cells, primary rat hippocampal neurons, and in whole-mouse brain sections, FGAMS immunoreactivity was distributed within the neuronal cytoplasm. FGAMS immunolabeling in vitro demonstrated extensive distribution throughout neuronal processes. To investigate potential changes in FGAMS expression and localization following viral infection, we infected cells with the human pathogen herpes simplex virus 1. In infected fibroblasts, FGAMS immunolabeling shifted from a diffuse cytoplasmic location to a mainly perinuclear localization by 12 h post-infection. In contrast, in infected neurons, FGAMS localization showed no discernable changes in the localization of FGAMS immunoreactivity. There were no changes in total FGAMS protein levels in either cell type. Together, these data provide insight into potential purine biosynthetic mechanisms utilized within neurons during homeostasis as well as viral infection. (Figure presented.). Cover Image for this Issue: doi: 10.1111/jnc.14169.
AB - Purines are metabolic building blocks essential for all living organisms on earth. De novo purine biosynthesis occurs in the brain and appears to play important roles in neural development. Phosphoribosyl formylglycinamidine synthase (FGAMS, also known as PFAS or FGARAT), a core enzyme involved in the de novo synthesis of purines, may play alternative roles in viral pathogenesis. To date, no thorough investigation of the endogenous expression and localization of de novo purine biosynthetic enzymes has been conducted in human neurons or in virally infected cells. In this study, we characterized expression of FGAMS using multiple neuronal models. In differentiated human SH-SY5Y neuroblastoma cells, primary rat hippocampal neurons, and in whole-mouse brain sections, FGAMS immunoreactivity was distributed within the neuronal cytoplasm. FGAMS immunolabeling in vitro demonstrated extensive distribution throughout neuronal processes. To investigate potential changes in FGAMS expression and localization following viral infection, we infected cells with the human pathogen herpes simplex virus 1. In infected fibroblasts, FGAMS immunolabeling shifted from a diffuse cytoplasmic location to a mainly perinuclear localization by 12 h post-infection. In contrast, in infected neurons, FGAMS localization showed no discernable changes in the localization of FGAMS immunoreactivity. There were no changes in total FGAMS protein levels in either cell type. Together, these data provide insight into potential purine biosynthetic mechanisms utilized within neurons during homeostasis as well as viral infection. (Figure presented.). Cover Image for this Issue: doi: 10.1111/jnc.14169.
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U2 - 10.1111/jnc.14304
DO - 10.1111/jnc.14304
M3 - Article
C2 - 29337348
AN - SCOPUS:85044382738
VL - 144
SP - 723
EP - 735
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
SN - 0022-3042
IS - 6
ER -