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JA Kink, ME Maley, RR Preston, KY Ling, MA Wallen-Friedman, Y Saimi & C Kung

Mutations in Paramecium CaM Indicate Functional Differences b/w the C-Terminal & N- Terminal Lobes In Vivo. JA Kink, ME Maley, RR Preston, KY Ling, MA Wallen-Friedman, Y Saimi & C Kung. Ashim Malhotra October 27, 2003. Lay Out. Background & Introduction CaM Structure: Rasmol

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JA Kink, ME Maley, RR Preston, KY Ling, MA Wallen-Friedman, Y Saimi & C Kung

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  1. Mutations in Paramecium CaM Indicate Functional Differences b/w the C-Terminal & N- Terminal Lobes In Vivo

  2. JA Kink, ME Maley, RR Preston, KY Ling, MA Wallen-Friedman, Y Saimi & C Kung Ashim Malhotra October 27, 2003

  3. Lay Out • Background & Introduction • CaM Structure: Rasmol • Experimentations • Conclusions • Philosophy & Trends

  4. Calmodulin • 148 amino acid long protein with 4 calcium ion binding pockets-I, II, III, IV. • III & IV in C-terminal loop & I & II in N-terminal loop. • Mediates many Ca signaling pathways. • Ca ion binding causes a “closed” conformation with hydrophobic domains inaccessible to targets.

  5. Calmodulin Structure

  6. www.angelfire.com/d20/bio_lab

  7. Paramecium Movement http://micro.magnet.fsu.edu/moviegallery/pondscum/protozoa/paramecium/

  8. Immunoblots of Mutant & wt CaMs

  9. Nucleotide Sequence of P. tetraurelia CaM gene

  10. Analysis of Paramecium CaM gene Northern Blot Southern Blot

  11. Amino Acid Sequence of wt & Mutant CaM genes

  12. Effect of CaM Mutations on Na+Behavior & Ca2+ Currents

  13. Ca2+ -Dependent Na+ Current & Ca2+ - Dependent K+ Current of wt, Cam3& Cam12Strains of P. teraurelia

  14. CaM Electrophoretic Mobilities Cosegregate w/ Behavioral Phenotypes among Autogamous F2 Segregants

  15. Conclusions • CaM mutation may not be lethal, instead it may cause behavioral/functional mutations. • In P. tetraurelia, behavior may be directly influenced by genetic mutations- the genetic dissection method for studying mutational effects. • The “over-reactor” mutants carry mutations in amino terminal half of CaM gene.

  16. Conclusions (contd…) • The “under-reactor” mutants carry mutations in C-terminal half of the gene. • The “under-reactors” show low inward Ca2+ -Dependent Na+ Currents. • The “over-reactors” show low Ca2+ - Dependent K+ Currents.

  17. The Philosophy • Functional or behavioral responses have a genetic origin ~ anthropological approaches for studying human development, particularly of intelligence. • A molecular phenomenology sited as causative link in an evolutionary process.

  18. Thank You

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