Charles K. Singleton, Ph.D.


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Faculty Appointments
Professor of Biological Sciences
Ph.D., Biochemistry, Purdue University, West Lafayette, IndianaB.S., University of Georgia, Athens, Georgia
Office Address
1210C Biological Sciences/MRB III
VU Station B 351634
Nashville, TN 37235-1634
Research Description
We investigate a variety of regulatory phenomena related to various aspects of development in Dictyostelium. This includes studies on sensory histidine kinases, trans-sensor ammonia transporters, and eIF2-alpha kinases in relation to their role in regulating aspects of multicellular development. The common theme is how communication amoung cells and between cells and their environment is accomplished and implemented to allow coordinated morphological development and cell differentiation. Our studies include the mechamisms underlying the regulation of the timing of development, the choice between alternative developmental pathways, the coordination of differentiation and morphogenesis, and the transition from growth to multicellular development. Our main approaches are genetic, molecular genetic, and biochemical.
Research Keywords
Cell-cell and cell-environment signaling during development.
Singleton CK, Xiong Y, Kirsten JH, Pendleton KP. EIF2a kinases regulate development through the BzpR transcription factor in Dictyostelium discoideum. PLoS ONE [print-electronic]. 2012; 7(3): e32500. PMID: 22403666, PMCID: PMC3293825, PII: PONE-D-11-21579, DOI: 10.1371/journal.pone.0032500, ISSN: 1932-6203.

Bowman RL, Xiong Y, Kirsten JH, Singleton CK. EIF2a kinases control chalone production in Dictyostelium discoideum. Eukaryotic Cell [print-electronic]. 2011 Apr; 10(4): 494-501. PMID: 21278229, PMCID: PMC3127634, PII: EC.00270-10, DOI: 10.1128/EC.00270-10, ISSN: 1535-9786.

Kirsten JH, Xiong Y, Davis CT, Singleton CK. Subcellular localization of ammonium transporters in Dictyostelium discoideum. BMC Cell Biol. 2008; 9: 71. PMID: 19108721, PMCID: PMC2653498, PII: 1471-2121-9-71, DOI: 10.1186/1471-2121-9-71, ISSN: 1471-2121.

Rai M, Xiong Y, Singleton CK. Disruption of the ifkA and ifkB genes results in altered cell adhesion, morphological defects and a propensity to form pre-stalk O cells during development of Dictyostelium. Differentiation. 2006 Dec; 74(9-10): 583-95. PMID: 17177855, PII: S0301-4681(09)60245-0, DOI: 10.1111/j.1432-0436.2006.00085.x, ISSN: 0301-4681.

Singleton CK, Kirsten JH, Dinsmore CJ. Function of ammonium transporter A in the initiation of culmination of development in Dictyostelium discoideum. Eukaryotic Cell. 2006 Jul; 5(7): 991-6. PMID: 16835443, PMCID: PMC1489295, PII: 5/7/991, DOI: 10.1128/EC.00058-06, ISSN: 1535-9778.

Kirsten JH, Xiong Y, Dunbar AJ, Rai M, Singleton CK. Ammonium transporter C of Dictyostelium discoideum is required for correct prestalk gene expression and for regulating the choice between slug migration and culmination. Dev. Biol [print-electronic]. 2005 Nov 11/1/2005; 287(1): 146-56. PMID: 16188250, PII: S0012-1606(05)00570-1, DOI: 10.1016/j.ydbio.2005.08.043, ISSN: 0012-1606.

Peter R. Martin, Charles K. Singleton, and Susanne Hiller-Sturmh?fel. The Role of Thiamine Deficiency in Alcoholic Brain Disease. Alcohol Research & Health. Alcohol Research & Health. 2004; 27: 134-42.

Follstaedt SC, Kirsten JH, Singleton CK. Temporal and spatial expression of ammonium transporter genes during growth and development of Dictyostelium discoideum. Differentiation. 2003 Dec; 71(9-10): 557-66. PMID: 14686953, PII: S0301-4681(09)60537-5, DOI: 10.1111/j.1432-0436.2003.07109002.x, ISSN: 0301-4681.

Fang R, Xiong Y, Singleton CK. IfkA, a presumptive eIF2 alpha kinase of Dictyostelium, is required for proper timing of aggregation and regulation of mound size. BMC Dev. Biol [print-electronic]. 2003 Apr 4/9/2003; 3: 3. PMID: 12697064, PMCID: PMC154100, ISSN: 1471-213X.

Martin PR, Singleton CK, Hiller-Sturmhöfel S. The role of thiamine deficiency in alcoholic brain disease. Alcohol Res Health. 2003; 27(2): 134-42. PMID: 15303623, ISSN: 1535-7414.

Song Q, Singleton CK. Mitochondria from cultured cells derived from normal and thiamine-responsive megaloblastic anemia individuals efficiently import thiamine diphosphate. BMC Biochem. 2002 Apr 4/25/2002; 3: 8. PMID: 12014993, PMCID: PMC111190, ISSN: 1471-2091.

Singleton CK, Martin PR. Molecular mechanisms of thiamine utilization. Curr. Mol. Med. 2001 May; 1(2): 197-207. PMID: 11899071, ISSN: 1566-5240.

Singleton, C. K. Transketolase. Encyclopedia of Molecular Medicine. 2001; 3224-322.

Wang, J. J-L., Hua, Z., Fentress, H. M., and Singleton, C. K. JNK1 is inactivated during thiamine deficiency-induced apoptosis in human neruoblastoma cells. 2000.

Ragan, P.W., Singleton, C.K., and Martin, P.R. Brain injury associated with chronic alcoholism. CNS Spectrums. 1999; 4: 66-87.

Singleton CK, Zinda MJ, Mykytka B, Yang P. The histidine kinase dhkC regulates the choice between migrating slugs and terminal differentiation in Dictyostelium discoideum. Dev. Biol. 1998 Nov 11/15/1998; 203(2): 345-57. PMID: 9808785, PII: S0012-1606(98)99049-2, DOI: 10.1006/dbio.1998.9049, ISSN: 0012-1606.

Zinda MJ, Singleton CK. The hybrid histidine kinase dhkB regulates spore germination in Dictyostelium discoideum. Dev. Biol. 1998 Apr 4/15/1998; 196(2): 171-83. PMID: 9576830, PII: S0012-1606(98)98854-6, DOI: 10.1006/dbio.1998.8854, ISSN: 0012-1606.

Pekovich SR, Martin PR, Singleton CK. Thiamine deficiency decreases steady-state transketolase and pyruvate dehydrogenase but not alpha-ketoglutarate dehydrogenase mRNA levels in three human cell types. J. Nutr. 1998 Apr; 128(4): 683-7. PMID: 9521628, ISSN: 0022-3166.

Pekovich, S. R., Martin, P. R., Poggi, V. and Singleton, C. K. Sensitivity to thiamine deficiency in cultered human cells is dependent on cell type and is enhanced in cells from thiamine- responsive megaloblastic anemia patients. 1998.

Singleton CK. Identification and characterization of the thiamine transporter gene of Saccharomyces cerevisiae. Gene. 1997 Oct 10/15/1997; 199(1-2): 111-21. PMID: 9358046, PII: S0378-1119(97)00354-5, ISSN: 0378-1119.

Wang JJ, Martin PR, Singleton CK. Aspartate 155 of human transketolase is essential for thiamine diphosphate-magnesium binding, and cofactor binding is required for dimer formation. Biochim. Biophys. Acta. 1997 Sep 9/5/1997; 1341(2): 165-72. PMID: 9357955, PII: S0167-4838(97)00067-8, ISSN: 0006-3002.

Wang JJ, Martin PR, Singleton CK. A transketolase assembly defect in a Wernicke-Korsakoff syndrome patient. Alcohol. Clin. Exp. Res. 1997 Jun; 21(4): 576-80. PMID: 9194907, PII: 00000374-199706000-00002, ISSN: 0145-6008.

Singleton CK, Wang JJ, Shan L, Martin PR. Conserved residues are functionally distinct within transketolases of different species. Biochemistry. 1996 Dec 12/10/1996; 35(49): 15865-9. PMID: 8961951, PII: bi9616920, DOI: 10.1021/bi9616920, ISSN: 0006-2960.

Pekovich SR, Martin PR, Singleton CK. Thiamine pyrophosphate-requiring enzymes are altered during pyrithiamine-induced thiamine deficiency in cultured human lymphoblasts. J. Nutr. 1996 Jul; 126(7): 1791-8. PMID: 8683340, ISSN: 0022-3166.

Wang, J.-L., Martin, P. R., and Singleton, C. K. Aspartate 155 of human transketolase is essential for thiamine pyrophosphate-magnesium binding, and cofactor binding is required for dimer formation. Biochem. et. Biophys. Acta. 1996; 1341: 165-72.

Martin PR, McCool BA, Singleton CK. Molecular genetics of transketolase in the pathogenesis of the Wernicke-Korsakoff syndrome. Metab Brain Dis. 1995 Mar; 10(1): 45-55. PMID: 7596328, ISSN: 0885-7490.

Singleton CK, Pekovich SR, McCool BA, Martin PR. The thiamine-dependent hysteretic behavior of human transketolase: implications for thiamine deficiency. J. Nutr. 1995 Feb; 125(2): 189-94. PMID: 7861245, ISSN: 0022-3166.

Martin PR, Pekovich SR, McCool BA, Whetsell WO, Singleton CK. Thiamine utilization in the pathogenesis of alcohol-induced brain damage. Alcohol Alcohol Suppl. 1994; 2: 273-9. PMID: 8974347, ISSN: 1358-6173.

Ken R, Singleton CK. Redundant regulatory elements account for the developmental control of a ribosomal protein gene of Dictyostelium discoideum. Differentiation. 1994 Jan; 55(2): 97-103. PMID: 8143933, ISSN: 0301-4681.

McPherson CE, Singleton CK. Nutrient-responsive promoter elements of the V4 gene of Dictyostelium discoideum. J. Mol. Biol. 1993 Jul 7/20/1993; 232(2): 386-96. PMID: 8345518, PII: S0022-2836(83)71398-7, DOI: 10.1006/jmbi.1993.1398, ISSN: 0022-2836.

Martin PR, McCool BA, Singleton CK. Genetic sensitivity to thiamine deficiency and development of alcoholic organic brain disease. Alcohol. Clin. Exp. Res. 1993 Feb; 17(1): 31-7. PMID: 8452206, ISSN: 0145-6008.

McCool BA, Plonk SG, Martin PR, Singleton CK. Cloning of human transketolase cDNAs and comparison of the nucleotide sequence of the coding region in Wernicke-Korsakoff and non-Wernicke-Korsakoff individuals. J. Biol. Chem. 1993 Jan 1/15/1993; 268(2): 1397-404. PMID: 8419340, ISSN: 0021-9258.

Martin, P.R., McCool, B.A., and Singleton, C.K. Genetic sensitivity to thiamine deficiency and development of alcoholic organic brain disease. Alcoholism: Clin. & Exp. Res. 1993; 17: 31-7.

McCool, B. A., Plonk, S. G., Martin, P. R., and Singleton, C. K. Cloning of human transketolase and comparison of the nucleotide sequence of the coding region in Wernicke-Korsakoff and non-Wernicke-Korsakoff individuals. J. Biol. Chem. 1993; 268: 1397-404.

McPherson CE, Singleton CK. V4, a gene required for the transition from growth to development in Dictyostelium discoideum. Dev. Biol. 1992 Apr; 150(2): 231-42. PMID: 1312963, PII: 0012-1606(92)90238-C, ISSN: 0012-1606.

Singleton CK, Delude RL, Ken R, Manning SS, McPherson CE. Structure, expression, and regulation of members of the developmentally controlled V and H gene classes from Dictyostelium. Dev. Genet. 1991; 12(1-2): 88-97. PMID: 2049884, DOI: 10.1002/dvg.1020120116, ISSN: 0192-253X.

Singleton CK, Manning SS, Ken R. Primary structure and regulation of vegetative specific genes of Dictyostelium discoideum. Nucleic Acids Res. 1989 Dec 12/11/1989; 17(23): 9679-92. PMID: 2602140, PMCID: PMC335206, ISSN: 0305-1048.

Singleton CK. Nucleotide sequence of V1, a ribosomal protein gene from Dictyostelium discoideum. Nucleic Acids Res. 1989 Oct 10/11/1989; 17(19): 7989. PMID: 2678007, PMCID: PMC334908, ISSN: 0305-1048.

Singleton CK, Gregoli PA, Manning SS, Northington SJ. Characterization of genes which are transiently expressed during the preaggregative phase of development of Dictyostelium discoideum. Dev. Biol. 1988 Sep; 129(1): 140-6. PMID: 2842207, PII: 0012-1606(88)90168-6, ISSN: 0012-1606.

Singleton CK, Manning SS, Feng Y. Effect of protein synthesis inhibition on gene expression during early development of Dictyostelium discoideum. Mol. Cell. Biol. 1988 Jan; 8(1): 10-6. PMID: 3336353, PMCID: PMC363071, ISSN: 0270-7306.

Singleton CK, McPherson CE, Manning SS. Deactivation of gene expression upon the onset of development in dictyostelium discoideum. Dev. Genet. 1988; 9(4-5): 327-35. PMID: 3243025, DOI: 10.1002/dvg.1020090414, ISSN: 0192-253X.

Singleton CK. PPAV, a promoter analysis vector for Dictyostelium discoideum. Nucleic Acids Res. 1987 Nov 11/25/1987; 15(22): 9610. PMID: 2825142, PMCID: PMC306502, ISSN: 0305-1048.

Singleton, C.K., Delude, R.L., and McPherson, C.E. Characterization of genes which are deactivated upon the onset of development of Dictyostelium discoideum. Dev. Biol. 1987; 119: 433-41.

Singleton, C.K. Left-handed helical DNA in solution. Topics in Nucleic Acids Structure 3, (S. Neidle, ed.).Macmillan Publishers LTD, Houndsmill, UK. 1987; 163-92.

Singleton, C.K. Structural heterogeneity of duplex DNA. Cell Proliferation: Recent Advances, vol. I (A.L. Boynton and H.L. Leffert, eds.). Academic Press, Inc., New York, NY. 1985; 417-47.

Singleton, C.K., Kilpatrick, M.W., and Wells, R.D. S1 nuclease recognizes DNA conformational junctions between left-handed helical (dT-dG)n .(dC-dA)n and contiguous right-handed sequences. J.Biol. Chem. 1984; 259: 1963-7.

Singleton, C.K. Effects of salts, temperature, and stem length on supercoil induced formation of cruciforms. J. Biol. Chem. 1983; 258: 7661-8.

Singleton, C.K., Klysik, J., and Wells, R.D. Conformational flexibility of junctions between contiguous B and Z DNA in supercoiled plasmids. Proc. Natl. Acad. Sci. USA. 1983; 80: 2447-51.

Singleton, C.K. and Wells, R.D. The facile generation of covalently closed, circular DNAs of defined negative superhelical densities. Anal. Biochem. 1982; 122: 253-7.

Singleton, C.K., Klysik, J., Stirdivant, S.M., and Wells, R.D. Left-handed Z-DNA is induced by supercoiling under physiological ionic conditions. Nature. 1982; 299: 312 316.