Source: UNIVERSITY OF NEBRASKA submitted to
THE ESSENTIAL ROLE OF BIOTIN IN CELL PROLIFERATION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
0189356
Grant No.
(N/A)
Project No.
NEB-91-053
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jul 1, 2001
Project End Date
Jun 30, 2006
Grant Year
(N/A)
Project Director
Zempleni, J.
Recipient Organization
UNIVERSITY OF NEBRASKA
(N/A)
LINCOLN,NE 68583
Performing Department
NUTRITIONAL & HEALTH SCIENCES
Non Technical Summary
Biotin deficiency and overdose may adversely affect growth of human cells. The purpose of this study is to identify roles of the vitamin biotin in cellular growth, proliferation, and differentiation.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
70260101010100%
Goals / Objectives
The long-term goal of the proposed research is to identify roles of the vitamin biotin in cellular growth, proliferation, and differentiation. In the studies proposed here, we will use human lymphocytes as a model to address the following objectives: 1. To quantify biotin transporter mRNA and transport activity in lymphocytes at various phases of the cell cycle. This objective will test the hypothesis that proliferating lymphocytes increase the expression of the gene encoding the biotin transporter at specific phases of the cell cycle. 2. To quantify biotinidase mRNA, biotinidase activity, and biotinylation of histones at various phases of the cell cycle in lymphocytes from healthy adults and biotinidase-deficient patients. This objective will test four hypotheses: (i) Proliferating lymphocytes increase the expression of the gene encoding biotinidase compared to nonproliferating controls. (ii) As a consequence, cellular biotinidase activity increases. (iii) Proliferating lymphocytes utilize increased amounts of biotin to biotinylate histones and, thus, the degree of biotinylation increases. (iv) Biotinylation of histones is reduced in biotinidase-deficient patients. 3. To identify enzymes that mediate debiotinylation of histones. This objective will test the hypothesis that the release of free biotin from biotinylated histones is an enzyme-mediated process. We will seek to identify this enzyme(s). 4. To determine whether biotinylation of histones is involved in DNA repair. Covalent modification of histones by poly (ADP-ribosylation) is linked to DNA repair mechanisms (Boulikas et al., 1990). This objective will test the hypothesis that biotinylation of histones is also linked to DNA repair mechanisms.
Project Methods
Objective 1: Expression of the biotin transporter gene in lymphocytes from various phases of the cell cycle will be determined by quantitative PCR and Northern blot analysis. Biotin transport in lymphocytes at various phases of the cell cycle will be determined using radioactive biotin as marker. Objective 2: Expression of the biotinidase gene in lymphocytes from various phases of the cell cycle will be determined by quantitative PCR and Northern blot analysis. Activity of biotinidase will be measured using a colorimetric procedure. Biotinylation of histones will be measured using Western blot procedures. Objective 3: Enzymes will be purified based on their physico-chemical properties or by labeling and purification of enzymes using photobiotin. Enzymes will be identified by protein sequencing. Objective 4: DNA damage in lymphocytes will be induced by exposure to nitrosurea or UV light. Biotinylation of histones will be measured using Western blot procedures and by using radioactive biotin as a marker.

Progress 07/01/01 to 06/30/06

Outputs
We have identified human genes the expression of which depends on the dietary supply of the vitamin biotin. Importantly, we discovered multiple novel cell signaling pathways that mediate biotin-dependent gene expression. One of these pathways is binding of biotin to histones (DNA-binding proteins). We demonstrated that biotinylation of histones plays crucial roles in gene regulation, DNA repair, and cell division. We identified 10 novel biotinylation sites in human histones, and we generated unique tools (antibodies; knockdown models) to investigate roles of histone biotinylation in chromatin structure.

Impacts
We have generated novel analytical tools (antibodies) to investigate roles of histones in cell biology. A patent application is currently pending and our invention has been licensed to Upstae, Inc. We have identified novel pathways of cell signaling by biotin in human cells: transcription factors Sp1, Sp3, and NF-kB. We also demonstrated that biotin affects signaling by receptor tyrosine kinases. Finally, we discovered that that biotin is bound covalently to histones, affecting chroamtin structure, gene expression, and DNA repair.

Publications

  • Rodriguez-Melendez R, Zempleni J. Biotin deficiency enhances nuclear translocation of NF-κB in Jurkat cells. FASEB J 18:A171, 2004
  • Sarath G, Kobza K, Rueckert B, Camporeale G, Zempleni J, Haas E. Biotinylation of human histone H3 and interactions with biotinidase. FASEB J 18:A103, 2004
  • Manthey KC, Zempleni J. Oxidative folding of secretory proteins is impaired in riboflavin-deficient HepG2 cells, triggering cell stress. FASEB J 18:A171, 2004
  • Kothapalli N, Zempleni J. Biotinylation of histones depends on the cell cycle in NCI-H69 small cell lung cancer cells. FASEB J 19:A55, 2005
  • Camporeale G, Suzuki Y, Eissenberg JC, Zempleni J. Low abundance of wild-type biotinidase and holocarboxylase synthetase decreases biotinylation of histones. FASEB J 19:A55, 2005
  • Herring T, Zempleni J, Cuppett S. Genomic implications of oxidative stress in Caco-2 cells. FASEB J 19:A1210, 2005
  • Griffin JB, Camporeale G, Zempleni J. Cell senescence is associated with decreased biotinylation of histone H4 in IMR90 human fibroblasts. FASEB J 20:A610, 2006
  • Pacheco-Alvarez D, Solorzano-Vargas RS, Gonzalez-Noriega A, Michalak C, Zempleni J, Leon-Del-Rio A. Biotin availability regulates expression of the sodium-dependent multivitamin transporter and the rate of biotin uptake in HepG2 cells. Mol Genet Metab 85:301-307, 2005
  • Rodriguez-Melendez R, Griffin JB, Zempleni J. The expression of genes encoding ribosomal subunits and eukaryotic translation initiation factor 5A depends on biotin and bisnorbiotin in HepG2 cells. J Nutr Biochem 17:23-30, 2006
  • Griffin JB, Rodriguez-Melendez R, Dode L, Wuytack F, Zempleni J. Biotin supplementation decreases the expression of the SERCA3 gene (ATP2A3) in Jurkat cells, thus, triggering unfolded protein response. J Nutr Biochem 17:272-281, 2006
  • Chew YC, Camporeale G, Kothapalli N, Sarath G, Zempleni J. Lysine residues in N- and C-terminal regions of human histone H2A are targets for biotinylation by biotinidase. J Nutr Biochem 17:225-233, 2006
  • Manthey KC, Rodriguez-Melendez R, Hoi JT, Zempleni J. Riboflavin deficiency causes protein and DNA damage in HepG2 cells, triggering arrest in G1 phase of the cell cycle. J Nutr Biochem 17:250-256, 2006
  • Camporeale G, Giordano E, Rendina R, Zempleni J, Eissenberg JC. Drosophila holocarboxylase synthetase is a chromosomal protein required for normal histone biotinylation, gene transcription patterns, lifespan, and heat tolerance. J Nutr 136:2735-2742, 2006
  • Chew YC, Sarath G, Zempleni J. An avidin-based assay for histone debiotinylase activity in human cell nuclei. J Nutr Biochem (2007)
  • Camporeale G, Oommen AM, Griffin JB, Sarath G, Zempleni J. K12-biotinylated histone H4 marks heterochromatin in human lymphoblastoma cells (2007)
  • Gralla M, Camporeale G, Zempleni J. Holocarboxylase synthetase regulates expression of biotin transporters by chromatin remodeling events at the SMVT locus (2006, submitted)
  • Camporeale G, Zempleni J, Eissenberg JC. Susceptibility to heat stress and aberrant gene expression patterns in holocarboxylase synthetase-deficient Drosophila melanogaster are caused by decreased biotinylation of histones rather than decreased biotinylation of carboxylases (2006, submitted)
  • Camporeale G, Eissenberg JC, Giordano E, Zempleni J. Lifespan and resistance to heat stress depend on histone biotinylation in Drosophila melanogaster. FASEB J 20:A610, 2006
  • Chew YC, Raza AS, Sarath G, Zempleni J. Biotinylation of K8 and K12 co-occurs with acetylation and mono-methylation in human histone H4. FASEB J 20:A610, 2006
  • Sarath G, Kobza K, Chew YC, Johnson K, Zempleni J, Raza A. Histone biotinylation in germinating switchgrass (Panicum virgatum L.) seeds. (in press) Association of Biomolecular Resource Facilities 2006 Annual Meeting February 11-14, 2006 in Long Beach, CA
  • Wijeratne SSK, Zempleni J. Decreased histone biotinylation marks cells senescence (2007)
  • Chew YC, West JT, Zempleni J. K12 biotinylated histone H4 is enriched at human endogenous retrovirus promoter regions and may function in retroviral silencing (2007)
  • Hassan YI, Atkin A, Zempleni J. Protein-protein interactions of human holocarboxylase synthetase reveals potential association with zinc-finger proteins (2007)
  • Kirkland J, Zempleni J, Buckles LK, Christman JK. Vitamin-dependent modifications of chromatin: epigenetic events and genomic stability. In: Handbook of Vitamins. Rucker RB, Zempleni J, Suttie JW, McCormick DB (eds), 4th edition. Taylor and Francis, Inc., Boca Raton, FL (2007)
  • Camporeale G, Zempleni J. Biotin. In: Present Knowledge in Nutrition. Bowman, BA, Russell RM (eds.), 9th edition, vol. I. International Life Sciences Institute, Washington, DC, 2006: 314 - 326, 2006
  • Berdanier C, Zempleni J. Advanced Nutrition. Taylor and Francis (2008)
  • Hassan YI, Zempleni J. Epigenetic regulation of chromatin structure and gene function by biotin. J Nutr 136:1763-1765, 2006


Progress 10/01/04 to 09/30/05

Outputs
We have identified novel pathways by which the vitamin biotin affects gene expression and DNA repair. First, we have shown that the cellular abundance of transcription factors Sp1 and Sp3 depends on biotin. Increased abundance of Sp1 and Sp3 in response to biotin supplementation is associated with increased expression of the CYP1B1 gene, enhaning the risk for DNA damage. Likewise, increased abundance of Sp1 and Sp3 mediates decreased expression of cellular calcium transporters, impairing the cellular folding and secretion of proteins. Second, we have shown that the cellular activity of the transcription factor NF-kB increases in response to biotin deficiency. This mediates activation of anti-apoptotic genes. Biotin deficiency is associated with decreased efficacy of cancer chemotherapy. Third, we have demonstrated that the vitamin biotin is bound covalently to histones (DNA-binding proteins). Further, we have demonstrated that biotinylation of histones plays a role in the cellular repsonse to DNA breaks, in the maintenance of pericentromeric chromatin, cell cycle progression, and gene silencing.

Impacts
We have identified the following novel roles for the vitamin biotin in cell biology. 1. Biotin affects gene expression. Effects of biotin on gene expression are mediated by transcription factors such as Sp1/Sp3 and NF-kB. 2. Biotin is covalently bound to histones, affecting chromatin structure. Biotinylation of histones plays roles in gene expression and DNA repair.

Publications

  • Sarath G, Kobza K, Chew YC, Johnson K, Zempleni J, Raza A. 2006. Histone biotinylation in germinating switchgrass (Panicum virgatum L.) seeds (submitted).
  • Camporeale G, Chew YC, Kueh A, Sarath G, Zempleni J. 2006. Use of synthetic peptides for identifying biotinylation sites in human histones. In: Avidin-Biotin Technology in the Life Sciences, Methods in Molecular Biology series. McMahon RJ (ed.). Humana Press, Totowa, NJ (invited manuscript, submitted).
  • Zempleni J, Christman J, Kirkland J. 2006. Vitamin-dependent modifications of chromatin: epigenetic events and genomic stability. In: Handbook of Vitamins. Rucker RB, Zempleni J, Suttie JW, McCormick DB (eds), 4th edition. Marcel Dekker, Inc., New York, NY (invited manuscript in preparation).
  • Oommen AM, Griffin JB, Sarath G, Zempleni J. 2005. Roles for nutrients in epigenetic events. J Nutr Biochem 16:74-77.
  • Kothapalli N, Camporeale G, Kueh A, Chew YC, Oommen AM, Griffin JB, Zempleni J. 2005. Biological functions of biotinylated histones. J Nutr Biochem 16:446-448.
  • Kothapalli N, Sarath G, Zempleni J. 2005. Biotinylation of K12 in histone H4 decreases in response to DNA double strand breaks in human JAr choriocarcinoma cells. J Nutr 135:2337-2342.
  • Oommen AM, Griffin JB, Zempleni J. 2005. K12-biotinylated histone H4 marks heterochromatin in human lymphoblastoma cells (submitted).
  • Kothapalli N, Zempleni J. 2005. Biotinylation of histones depends on the cell cycle in NCI-H69 small cell lung cancer cells. FASEB J 19:A55.
  • Camporeale G, Suzuki Y, Eissenberg JC, Zempleni J. 2005. Low abundance of wild-type biotinidase and holocarboxylase synthetase decreases biotinylation of histones. FASEB J 19:A55.
  • Herring T, Zempleni J, Cuppett S. 2005. Genomic implications of oxidative stress in Caco-2 cells. FASEB J 19:A1210.
  • Kothapalli N, Chew YC, Zempleni J. 2005. Histone biotinyl transferase activity depends on p53 in HCT 116 colon cancer cells. Cancer Epidemiol Biomarkers Prev (in press).
  • Griffin JB, Camporeale G, Zempleni J. 2006. Cell senescence is associated with decreased biotinylation of histone H4 in IMR90 human fibroblasts. FASEB J (in press).
  • Camporeale G, Eissenberg JC, Giordano E, Zempleni J. 2006. Lifespan and resistance to heat stress depend on histone biotinylation in Drosophila melanogaster. FASEB J (in press).
  • Chew YC, Raza AS, Sarath G, Zempleni J. 2006. Biotinylation of K8 and K12 co-occurs with acetylation and mono-methylation in human histone H4. FASEB J (in press).
  • Rodriguez-Melendez R, Griffin JB, Zempleni J. 2005. The expression of genes encoding ribosomal subunits and eukaryotic translation initiation factor 5A depends on biotin and bisnorbiotin in HepG2 cells. J Nutr Biochem (in press).
  • Griffin JB, Zempleni J. 2005. Biotin deficiency stimulates survival pathways in human lymphoma cells exposed to antineoplastic drugs. J Nutr Biochem 16:96-103.
  • Griffin JB, Rodriguez-Melendez R, Dode L, Wuytack F, Zempleni J. 2005. Biotin supplementation decreases the expression of the SERCA3 gene (ATP2A3) in Jurkat cells, thus, triggering unfolded protein response. J Nutr Biochem (in press).
  • Kobza K, Camporeale G, Rueckert B, Kueh A, Griffin JB, Sarath G, Zempleni J. 2005. K4, K9, and K18 in human histone H3 are targets for biotinylation by biotinidase. FEBS J 272:4249-4259.
  • Rodriguez-Melendez R, Griffin JB, Sarath G, Zempleni J. 2005. High-throughput immunoblotting identifies biotin-dependent signaling proteins in HepG2 hepatocarcinoma cells. J Nutr 135:1659-1666.
  • Manthey KC, Chew YC, Zempleni J. 2005. Riboflavin deficiency impairs oxidative folding and secretion of apolipoprotein B-100 in HepG2 cells, triggering stress-response systems. J Nutr 135:978-982.
  • Werner R, Manthey KC, Griffin JB, Zempleni J. 2005. HepG2 cells develop signs of riboflavin deficiency within four days of culture in riboflavin-deficient medium. J Nutr Biochem 16:617-624.
  • Chew YC, Camporeale G, Kothapalli N, Sarath G, Zempleni J. 2005. Lysine residues in N- and C-terminal regions of human histone H2A are targets for biotinylation by biotinidase. J Nutr Biochem.
  • Manthey KC, Rodriguez-Melendez R, Hoi JT, Zempleni J. 2005. Riboflavin deficiency causes protein and DNA damage in HepG2 cells, triggering arrest in G1 phase of the cell cycle. J Nutr Biochem (in press).


Progress 10/01/03 to 09/29/04

Outputs
We have identified novel pathways by which the vitamin biotin affects gene expression and DNA repair. First, we have shown that the cellular abundance of transcription factors Sp1 and Sp3 depends on biotin. Biotin supplementation is associated with transcriptional activation of the Sp1-dependent gene coding for cytochrome P450 1B1 (CYP1B1). Activation of CYP1B1 is associated with increased frequency of DNA breaks in human cells, enhancing the risk for tumor initiation. Second, we have shown that biotin deficiency is associated with increased nuclear translocation of the transcription factor NF-kB. NF-kB is associated with transcriptional activation of anti-apoptotic genes. Potentially, biotin deficiency is associated with decreased efficacy of cancer chemotherapy. Third, we have developed a tool to identify biotinylation sites in histones (DNA-binding proteins). Using this tool, we have identified seven novel binding sites for biotin in histones. We demonstrated that biotinylation of histones might play a role in mediating DNA repair.

Impacts
We have identified the following novel roles for the vitamin biotin in cell biology. 1. Biotin affects gene expression. Effects of biotin on gene expression are mediated by transcription factors such as Sp1/Sp3 and NF-kB. 2. Biotin is covalently bound to histones, affecting chromatin structure. Biotinylation of histones plays roles in gene expression and DNA repair.

Publications

  • Landenberger A, Kabil H, Harshman LG, Zempleni J. 2004. Biotin deficiency decreases life span and fertility but increases stress resistance in Drosophila Melanogaster. J Nutr Biochem.
  • Wiedmann S, Rodriguez-Melendez R, Ortega-Cuellar D, Zempleni J. 2004. Clusters of biotin-dependent genes in human peripheral blood mononuclear cells. J Nutr Biochem 15:433-439.
  • Camporeale G, Shubert EE, Sarath G, Cerny R, Zempleni J. K8 and K12 are biotinylated in human histone H4. 2004. Eur J Biochem 271:2257-2263.
  • Rodriguez-Melendez R, Schwab LD, Zempleni J. 2004. Jurkat cells respond to biotin deficiency with increased nuclear translocation of NF-kB, mediating cell survival Int J Vitam Nutr Res 74:209-216.
  • Rodriguez-Melendez R, Griffin JB, Zempleni J. 2004. Biotin supplementation increases expression of the cytochrome P450 1B1 gene in Jurkat cells, increasing the occurrence of single-stranded DNA breaks. J Nutr 134:2222-2228.
  • Camporeale G, Shubert EE, Sarath G, Zempleni J. 2004. Lysine-8 and lysine-12 are biotinylated in human histone H4. FASEB J 18:A104.
  • Kothapalli N, Zempleni J. 2004. Double strand breaks of DNA decrease biotinylation of lysine-12 in histone H4 in JAr cells. FASEB J 18:A103-104.
  • Rodriguez-Melendez R, Zempleni J. 2004. Biotin deficiency enhances nuclear translocation of NF-kB in Jurkat cells. FASEB J 18:A171
  • Sarath G, Kobza K, Rueckert B, Camporeale G, Zempleni J, Haas E. 2004. Biotinylation of human histone H3 and interactions with biotinidase. FASEB J 18:A103.
  • Manthey KC, Zempleni J. 2004. Oxidative folding of secretory proteins is impaired in riboflavin-deficient HepG2 cells, triggering cell stress. FASEB J 18:A171.
  • Camporeale G, Kothapalli N, Sarath G, Zempleni J. 2005. Biotinylation of histones in human cells. In: Nutrients and Cell Signaling. Zempleni J. Dakshinamurti K (eds.). Marcel Dekker, New York, NY (in press).
  • Camporeale G, Rodriguez-Melendez R, Zempleni J. 2005. Pantothenic acid and biotin. In: Sports Nutrition: Vitamins and Trace Elements. Driskell JA, Wolinsky I (eds.). CRC Press, Boca Raton, FL (invited review, submitted)
  • Zempleni J. 2005. Uptake, Localization, and Noncarboxylase Roles for Biotin. Annu Rev Nutr (invited review, submitted).


Progress 10/01/02 to 09/30/03

Outputs
1. We have identified a new biotin transporter in human lymphoid cells: monocarboxylate transporter 1 (MCT1). Mutations in MCT1 may account for inborn errors in biotin transport in humans. 2. We have developed an analytical procedure to identify biotinylation sites in human histones. This procedure is based on using chemically synthesized peptides (fragments of histones) as substrates for enzymatic biotinylations. 3. We have identified two biotinylation sites in human histone H4: lysine-8 and lysine-12. Also, we have demonstrated that acetylation of lysines reduces biotinylation of neighboring lysines in histone H4. 4. We have generated a polyclonal antibody to histone H4, biotinylated at lysine-12. 5. The antineoplastic agent etoposide causes double-strand breaks in DNA. We have shown that treatment of human cells with etoposide causes a rapid decrease in biotinylation of lysine-12 in histone H4. 6. We quantified gene expression profiles in lymphocytes from healthy adults before and after supplementation with biotin. Patterns of gene expression in lymphocytes were quantified by DNA microarray and RT-PCR for 450 genes. Biotin supplementation increased expression of genes encoding interferon-g, interleukin-1b, and 3-methylcrotonyl-CoA carboxylase, and decreased expression of the gene encoding interleukin-4. These findings suggest that biotin affects expression of genes encoding cytokines, possibly affecting cell proliferation. 7. We have provided evidence that nuclear abundance of transcription factors Sp1 and Sp3 correlates with biotin status in human T cells; that biotin affects the transcriptional activity of genes encoding Sp1 and Sp3; and that reduced abundance of Sp1 and Sp3 in biotin-deficient cells is associated with decreased transcriptional activity of Sp1/Sp3-dependent genes. 8. Cells respond to various kinds of stress with increased nuclear translocation of NF-kB, mediating transcriptional activition of anti-apoptotic genes. We have shown that incubation of human T cells in biotin-deficient medium for 5 weeks is associated with increased nuclear translocation of NF-kB compared to biotin-sufficient controls. Nuclear translocation of NF-kB is associated with transcriptional activation of the anti-apoptotic Bfl-1 gene in biotin-deficient cells; activation of Bfl-1 mediates survival during phases of biotin deficiency. Consistent with this hypothesis, biotin-deficient cells survive exposure to serum starvation and tumor necrosis factor alpha more efficiently than biotin-supplemented controls; activity of the apoptotic enzyme caspase-3 is lower in biotin-deficient cells compared to controls. Nuclear translocation of NF-kB also activates the p53 gene, mediating decreased rates of cell proliferation in biotin-deficient cells compared to biotin-sufficient controls. 9. Most biotin catabolites have been considered metabolic waste in previous studies. Using synthetic biotin analogs we have now shown that synthetic biotin catabolites have biotin-like activities in human cells.

Impacts
We have identified the following novel roles for the vitamin biotin in cell biology. 1. Biotin affects gene expression. Effects of biotin on gene expression are mediated by transcription factors such as Sp1/Sp3 and NF-kB. 2. Biotin is covalently bound to histones, affecting chromatin structure. Biotinylation of histones plays roles in gene expression and DNA repair.

Publications

  • Wiedmann S, Eudy JD, Zempleni J. 2003. Biotin supplementation increases expression of genes encoding interferon-gamma, interleukin-1beta, and 3-methylcrotonyl-CoA carboxylase, and decreases expression of the gene encoding interleukin-4 in human peripheral blood mononuclear cells. J Nutr 133:716-719.
  • Camporeale G, Zempleni J. 2003. Oxidative folding of interleukin-2 is impaired in flavin-deficient Jurkat cells, causing intracellular accumulation of interleukin-2 and increased expression of stress response genes. J Nutr 133:668-672.
  • Crisp SERH, Camporeale G, White B, Toombs CF, Griffin JB, Said HM, Zempleni J. 2003. Biotin supply affects rates of cell proliferation, biotinylation of carboxylases and histones, and expression of the gene encoding the sodium-dependent multivitamin transporter in JAr choriocarcinoma cells. Eur J Nutr (in press).
  • Rodriguez-Melendez R, Lewis B, McMahon RJ, Zempleni J. 2003. Diaminobiotin and desthiobiotin have biotin-like activities in Jurkat cells. J Nutr 133:1259-1264.
  • Scheerger SB, Zempleni J. 2003. Expression of oncogenes depends on biotin in human small cell lung cancer cells NCI-H69. Int J Vitam Nutr Res (in press).
  • Daberkow RL, White BR, Cederberg RA, Griffin JB, Zempleni J. 2003. Monocarboxylate transporter 1 mediates biotin uptake in human peripheral blood mononuclear cells. J Nutr 133:2703-2706.
  • Griffin JB, Rodriguez-Melendez R, Zempleni J. 2003. The nuclear abundance of transcription factors Sp1 and Sp3 depends on biotin in Jurkat cells. J Nutr (in press).
  • Rodriguez-Melendez R, Schwab LD, Zempleni J. 2004. Jurkat cells respond to biotin deficiency ... (submitted)
  • Landenberger A, Kabil H, Harshman L, Zempleni J. 2004. Biotin deficiency decreases life span and fertility but increases stress resistance in Drosophila melanogaster (manuscript in preparation).
  • Camporeale G, Sarath G, Shubert EE, Zempleni J. 2004. Lysines 8 and 12 are biotinylated in human histone H4 (manuscript in preparation).
  • Camporeale G, Kothapalli N, Sarath G, Zempleni J. 2004. Biotinylation of histones in human cells. In: Zempleni J and Dakshinamurti K (eds.). Nutrients and Cell Signaling. Marcel Dekker, New York, NY (submitted).
  • Rodriguez-Melendez R, Zempleni J. 2003. Regulation of gene expression by biotin. J Nutr Biochem (in press).
  • Zempleni J, Camporeale G. 2003. Riboflavin deficiency impairs oxidative folding of interleukin-2, triggering unfolded protein response in Jurkat cells. FASEB J 17:A277.
  • Griffin JB, Crisp SERH, Camporeale G, White BR, Toombs CF, Said HM, Zempleni J. 2003. Biotin supply affects proliferation, biotinylation of proteins, and expression of biotin transporter genes in JAr cells. FASEB J 17:A720.
  • Rodriguez-Melendez R, Wiedmann S, Eudy JD, Zempleni J. 2003. Biotin supplementation affects gene expression in lymphocytes from healthy adults. FASEB J 17:A720.


Progress 10/01/01 to 09/30/02

Outputs
1. Biotin transporter studies: We have generated a polyclonal antibody to the human biotin transporter "SMVT". Using this antibody we have investigated biotin transport in rapidly proliferating human cells, e.g., immune cells and placental cells. Expression of SMVT increased early in the cell cycle, leading to increased cellular biotin uptake and increased biotinylation of carboxylases and histones. These findings are consistent with the hypothesis that rapidly proliferating human cells have an increased demand for biotin. 2. Immune function: Cellular biotin status affects expression of genes that play roles in immune function (cytokines and their receptors) in healthy adults and in cell lines. This finding suggests that biotin has beneficial effects regarding immune function. 3. Human placenta: We have demonstrated that biotin deficiency adversely affects proliferation rates of human placental cells, and secretion of hormones by these cells. Impaired function of placenta might account for some of the fetal malformations seen in biotin-deficient women. 4. Histones: We have demonstrated that histones are biotinylated in human cells, that debiotinylation of histones is mediated by the enzyme biotinidase, and that biotinylation of histones might play a role in preventing tumor initiation.

Impacts
1. We have shown that proliferating cells have an increased demand for biotin, as judged by increased biotin uptake, biotinylation of histones, and carboxylase activities in proliferating cells. Our studies will help to estimate biotin requirements of growing tissues/organs in vivo. 2. Our studies provided evidence that biotinylation of histones might play a role in cell proliferation, repair of damaged DNA, and apoptosis. These findings are consistent with an essential for biotin in the prevention of tumor initiation. 3. The enzyme biotinidase plays a role in regulating biotinylation stoichiometry of histones. In the inborn error "biotinidase deficiency" (prevalence 1/60,000), biotinidase activity is reduced to less than 30% of normal controls. Our studies will improve understanding of roles for biotinidase in cell proliferation and tumor initiation. 4. Biotin affects expression of numerous genes that play a role in immune function, consistent with beneficial effects of biotin in the immune system.

Publications

  • Stanley, J.S., Griffin, J.B., Zempleni, J. 2001. Biotinylation of histones in human cells: effects of cell proliferation. Eur. J. Biochem. 268:5424-5429.
  • Ballard, T.D., Wolff, J., Griffin, J.B., Stanley, J.S., van Calcar, S., Zempleni, J. 2002. Biotinidase catalyzes debiotinylation of histones. Eur. J. Nutr. 41:78-84.
  • Griffin, J.B., Stanley, J.S., Zempleni, J. 2002. Synthesis of a rabbit polyclonal antibody to the human sodium-dependent multivitamin transporter. Int. J. Vitam. Nutr. Res. 72:195-198.
  • Manthey, K.C., Griffin, J.B., Zempleni, J. 2002. Biotin supply affects expression of biotin transporters, biotinylation of carboxylases, and metabolism of interleukin-2 in Jurkat cells. J. Nutr. 132:887-892.
  • Stanley, J.S., Mock, D.M., Griffin, J.B., Zempleni, J. 2002. Biotin uptake into human peripheral blood mononuclear cells increases early in the cell cycle, increasing carboxylase activities. J. Nutr. 132:1854-1859.
  • Peters, D.M., Griffin, J.B., Stanley, J.S., Beck, M.M., Zempleni, J. 2002. Exposure to UV light causes increased biotinylation of histones in Jurkat cells. Am. J. Physiol. Cell Physiol. 283:878-884.
  • Rodriguez-Melendez, R., Camporeale, G., Griffin, J.B., Zempleni, J. 2002. Interleukin-2 receptor gamma-dependent endocytosis depends on biotin (in press).
  • Zempleni, J. Biotin. 2001. In: Present Knowledge in Nutrition. International Life Sciences Institute, Washington, DC, pp. 241-252.
  • Zempleni, J. 2003. Biotinylation of histones. In: Zempleni, J. and Daniel, H. (eds.) Molecular Nutrition. CABI Press, UK (in press)
  • Zempleni, J. 2003. Nutrient homeostasis in proliferating cells. In: Zempleni J and Daniel H. (eds.) Molecular Nutrition. CABI Press, UK (in press)
  • Zempleni, J., Manthey, K.C., Griffin, J.B. 2002. Biotin affects secretion secretion of interleukin-2 by Jurkat cells. FASEB J. 16:614.