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Anaerobiosis: a brief look at genes in human and yeast

Anaerobiosis: a brief look at genes in human and yeast. Michelle Mousel Patty Keating Erickson Vicher. Issues addressed will include:. What are the genes conserved between yeast and humans? How are these genes regulated? Why is this information valuable?. Phylogenetic Tree. Human.

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Anaerobiosis: a brief look at genes in human and yeast

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  1. Anaerobiosis: a brief look at genes in human and yeast Michelle Mousel Patty Keating Erickson Vicher

  2. Issues addressed will include: • What are the genes conserved between yeast and humans? • How are these genes regulated? • Why is this information valuable?

  3. Phylogenetic Tree Human Photosynthetic Bacteria Baker’s yeast Insects N. crassa Plants

  4. Aerobic Respiration C6H12O6 + 6 O2 + 38 ADP +38 Pi 6 CO2 + 44 H2O + 38 ATP Human & Yeast same pathway 19 times more efficient

  5. Anaerobic Respiration Humans: C6H12O6 + 2 ADP +2 Pi 2 Lactate + 2 H+ + 2 H20 + 2 ATP Yeast: C6H12O6 + 2 ADP +2 Pi 2 Ethanol + 2 CO2 + 2 H20 + 2 ATP

  6. Human/Yeast homologues involved in respiratory processes HUMAN YEASTFUNCTION Aerobic and anaerobic in yeast; promotes formation of first peptide bond in translation Superoxide dismutase; provides protection from oxygen toxicity ADP/ATP transporter in mitochondria involved in glucose repression cytochrome c oxidase Eif5-a TIF51A-B SOD SOD1 ANT2 AAC3 TUPLE1 TUP1 COX15 COX15

  7. GCG Pileup of human eif5-a vs. yeast tif51b

  8. GCG pileup of superoxide dismutase from human and yeast

  9. GCG bestfit of human TUPLE1 and yeast TUP1

  10. GCG pileup of human ANT2 vs. yeast AAC3 and ACC2

  11. Nucleotide sequence comparison of human ANT2 vs yeast AAC3 ADT/ATP transporter Due to complex intron exon structure, nucleotide homology appears lower than protein.

  12. Protein sequence homology shows conservation of the proteins involved in aerobic and anaerobic processes Most of the genes involved in respiration are regulated at a transcriptional level These regulators include:

  13. YEAST HUMAN ROX1-represses most known anaerobic genes consensus binding site:YYATTGTTCTC represses:HEM13, TIF51-b, ERG11, ANB1, COX5, AAC3 O2 and Heme- induces ROX1; represses: DAN1 HAP1-induces many anaerobic genes including: ERG11, TIF51-a, CYC7, HEM13, HAP2/3/4-induces: COX genes HIF1-hypoxia inducing factors activates many of the enymes in anaerobic glycolysis targets include: ALD-A, PGK1, enolase1, LDH-A, PFKL other genes that show HIF1 binding include: TH, VEGF, GLUT1 WHERE ARE THE OTHER REGULATORS?

  14. Regulatory consensus sites Activator sites: Hem13UAS (GGAGTCG)2 ERG11 GCCCGGGAATTACCGGGGGC TTACGTACTCGCATGTATT HIF1 Regulates ALD-A, PGK1, ENOLASE1, LDH-A, PFK L, PGD1 Binds TH VEGF, GLUT1 TACGTGCT GRBOX ANT2 ATGATT TIF51B GTTCGTTGCCT Repressor sites: Rox1: HEM13, TIF51B YYYATTGTTCTC ERG11 AAACGAUYAAACGAA AAACGAAACGAAACGAAACTAA ANB1 AAGGAACGAACAA COX5B AACGAACGA Consensus YYATTGTTCTC CATTGTT CCATTGTTC

  15. Future projects: • Look for repressors of human anaerobiosis, possibly like ROX1, as the ANT2 regulatory sequence is similar to the ROX1 consensus site; need more sequences from human sequencing projects for this as well as the next one • Look for yeast inducers of anaerobic genes that may be like HIF1 genes in humans • Further studies of expressing yeast proteins in mammalian systems; and human proteins in yeast systems

  16. Why Yeast? • Easy to manipulate • The entire genome is sequenced • Can make an entirely aerobic or anaerobic system

  17. Why study aerobic and anaerobic respiration? • Initial topic of interest • Lactate buildup during exercise • athletes and athletics • release of lysosomes, cell death, soreness • WWW Search for lactate buildup • Yahoo - 12393 hits • Excite - 19858 hits

  18. Some examples • Devices used to monitor lactate levels • Accusport Lactate Analyzer • Endurex Excel dietary supplement • Horses also addressed • Vita-Flex Lactanase

  19. Secondary topic of interest • Diseases • Cancer • HIV • Influenza all anaerobic

  20. Our purpose Elucidation Data mining

  21. That's all Folks!!

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