Research Interests:Aging appears to be the progressive loss of function through the deterioration of biological systems that control repair and maintenance. A multitude of factors are known to change with age, such as methylation patterns, enzyme activities, and production of inflammatory molecules. Clearly there are many changes in gene expression and changes in genes with age, but I believe primary authority over regulation of the rate of aging resides in the innate sequence of the genome. DNA encodes an organism’s biological processes, including its own repair and protection biomachinery. The rate of repair and protection and the rate of damage (whether endogenously or exogenously inflicted) together dictate the rate of deterioration of an organism. My long-standing interest is the aging process, and my goal at the Institute for Genome Sciences is to employ the power of modern genomic technology to elucidate genes associated with longevity and the genetic changes that contributed to the evolution of human longevity. There are widely divergent lifespans among primate species, yet it appears that genes that modulate aging have been conserved in sequence and function among divergent species over millions of years. Neoteny has been proposed as a mechanism that contributed to the rise of many human traits, including increased life span as compared to other primates. Comparative genomics will reveal the biosecrets that allow humans to age more slowly than other primates. An analysis that determines to what extent the genetic variants that modulate longevity are conserved across different species could have useful biomedical applications. Although aging has been slowed and healthy lifespan prolonged in many different animal models, much remains to be understood. Do the longer-lived species have a more stable genetic apparatus? Superior repair and protection mechanisms of cellular constituents? The comparative genomics approach will allow us to decode which regions of the genome are responsible for the preservation and extension of health in longer-lived species.
Lab Techniques and Equipment:• PCR. qPCR. Primer and Probe design. Gene synthesis. Cloning.
• DNA purification: mammalian genomic, phage, bacterial, plasmid.
• DNA modification: nucleases, endonucleases, ligase, kinase, phosphatase.
• Gel electrophoresis of DNA: agarose and polyacrylamide.
• FIGE (Field Inversion Gel Electrophoresis).
• TAFE (Transverse Alternating Field Electrophoresis).
• Site Directed Mutagenesis.
• Homologous recombination.
• Southern Blots.
• Library Construction.
• Mammalian tissue culture maintenance. Sterile technique.
• Mammalian clonal cell lines: Initiation, growth, splitting, cryogenic storage.
• Hybridoma cell line creation for production of monoclonal antibodies.
• Cell fractionation: isolation of microsomes, SER, RER.
• Isolation of cell nuclei, extraction of chromatin (rodent and primate tissue).
• Protein Purification Techniques.
• Column chromatography: Ion-exchange, affinity, and exclusion.
• Cyanogen bromide activation of sepharose 4B affinity gel.
• ELISA. Protein Detection and Quantitation. Enzyme assays.
• Peptide conjugation.
• IgG, IgM immunodetection; immunodiffusion test.
• SDS Polyacrylamide gel electrophoresis. Tricine gels.
• Western Blots.
• Fluorescence anisotropy measurements by steady state fluorimeter.
• Bacteria cells: Growth, induction, cryogenic storage.
• Purification of Lambda phage, T4 phage, M13.
• Phage T4 Expression-Packaging-Processing System to package and produce proteins.
• Production of proteins: by IPTG induction, heat-induction.
• Construction and production of chimera proteins.
• Phage Display.
Mills, R.E., Pittard, W.S., Mullaney, J.M., Farooq, U., Creasy, T.H., Mahurkar, A.A., Kemeza, D.M., Strassler, D.S., Ponting, C.P., Webber, C., and Devine, S.E. 2011. Natural genetic variation caused by small insertions and deletions in the human genome. Genome Res. 2011 Apr 25. [Epub ahead of print] PMID: 21460062
Mullaney, J.M., Mills, R.E., Pittard, W.S., and Devine, S.E. 2010. Small insertions and deletions (INDELs) in human genomes. Hum Mol Genet. Oct 15;19(R2):R131-6. PMID: 20858594
Baumann, R.G., Mullaney, J.M. and Black, L.W. 2006. Portal fusion protein constraints on function in DNA packaging of bacteriophage T4. Mol Microbiol. Jul;61(1):16-32. PMID: 16824092
Mullaney, J.M., Thompson, R.B., Gryczynski, Z., and Black, L.W. 2000. Green fluorescent protein as a probe of rotational mobility within bacteriophage T4. J Virol Methods. 2000 Jul;88(1):35-40. PMID: 10921840
Mullaney, J.M. and Black, L.W. 1998. GFP:HIV-1 protease production and packaging with a T4 phage expression-packaging-processing system. Biotechniques. Dec;25(6):1008-12. PMID: 9863054
Mullaney, J.M. and Black, L.W. 1998. Activity of foreign proteins targeted within the bacteriophage T4 head and prohead: implications for packaged DNA structure. J Mol Biol. Nov 13;283(5):913-29. PMID: 9799633
Mullaney, J.M., and Black, L.W. 1996. Capsid targeting sequence targets foreign proteins into bacteriophage T4 and permits proteolytic processing. J Mol Biol. Aug 23;261(3):372-85. PMID: 8780780
Hong, Y.R., Mullaney, J.M., and Black, L.W. 1995. Protection from proteolysis using a T4::T7-RNAP phage expression-packaging-processing system. Gene. Aug 30;162(1):5-11.PMID: 7557416
Ghosh, T.K., Mullaney, J.M., Tarazi, F.I., and Gill, D.L. 1989. GTP-activated communication between distinct inositol 1,4,5-trisphosphate-sensitive and -insensitive calcium pools. Nature. Jul 20;340(6230):236-9. PMID: 2787892
Gill, D.L., Ghosh, T.K., Mullaney, J.M. 1989. Calcium signalling mechanisms in endoplasmic reticulum activated by inositol 1,4,5-trisphosphate and GTP. Cell Calcium. Jul;10(5):363-74. Review. PMID: 2670240
Gill, D.L., Mullaney, J.M., and Ghosh, T.K. 1988. Intracellular calcium translocation: mechanism of activation by guanine nucleotides and inositol phosphates. J Exp Biol. Sep;139:105-33. Review. PMID: 3062118
Ghosh, T.K., Eis, P.S., Mullaney, J.M., Ebert, C.L., and Gill, D.L. 1988. Competitive, reversible, and potent antagonism of inositol 1,4,5-trisphosphate-activated calcium release by heparin. J Biol Chem. Aug 15;263(23):11075-9. PMID: 3136153
Mullaney, J.M., Yu, M., Ghosh, T.K., and Gill, D.L. 1988. Calcium entry into the inositol 1,4,5-trisphosphate-releasable calcium pool is mediated by a GTP-regulatory mechanism. Proc Natl Acad Sci U S A. Apr;85(8):2499-503. PMID: 3357878
Mullaney, J.M., Chueh, S.H., Ghosh, T.K., and Gill, D.L. 1987. Intracellular calcium uptake activated by GTP. Evidence for a possible guanine nucleotide-induced transmembrane conveyance of intracellular calcium. J Biol Chem. Oct 5;262(28):13865-72. PMID: 3654640
Chueh, S.H., Mullaney, J.M., Ghosh, T.K., Zachary, A.L., and Gill, D.L. 1987. GTP- and inositol 1,4,5-trisphosphate-activated intracellular calcium movements in neuronal and smooth muscle cell lines. J Biol Chem. Oct 5;262(28):13857-64. PMID: 3498720
Tolmasoff, J.M., Ono, T., and Cutler, R.G. 1980. (J.M. Mullaney's Maiden Name: Tolmasoff) Superoxide dismutase: Correlation with lifespan and specific metabolic rate in primate species. Proc Natl Acad Sci USA. May;77(5):2777-81. PMID: 6771758
Moment, G.B., Tolmasoff, J.M., and Cutler, R.G. 1980. (J.M. Mullaney's Maiden Name: Tolmasoff) Superoxide dismutase, thermal respiratory acclimation, and growth in an earthworm, Eisenia foetida. Growth Sep;44(3):230-4.
Links of Interest:American Aging Association
American Federation for Aging Research
Gerontology Research Group
Human Aging Genomic Resources
National Institute on Aging
Supercentenarian Research Foundation
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