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Excision of Intron Sequences

Excision of Intron Sequences

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Excision of Intron Sequences

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  1. Excision of Intron Sequences

  2. Splicing • Removal of introns must be very precise. • Conserved sequences for removal of the introns of nuclear mRNA genes are minimal. • Dinucleotide sequences at the 5’ and 3’ ends of introns. • An A residue about 30 nucleotides upstream from the 3’ splice site is needed for lariat formation.

  3. Types of Intron Excision • The introns of tRNA precursors are excised by precise endonucleolytic cleavage and ligation reactions catalyzed by special splicing endonuclease and ligase activities. • The introns of nuclear pre-mRNA (hnRNA) transcripts are spliced out in two-step reactions carried out by spliceosomes.

  4. The Spliceosome • Five snRNAs: U1, U2, U4, U5, and U6 • Some snRNAs associate with proteins to form snRNAs (small nuclear ribonucleoproteins)

  5. What are Logo plots?

  6. Logo for a) Splice acceptor b) Splice Donor c) Initiator Met

  7. AG/GT CAG/NT exon 1 intron 1 exon 2 4 3 2 1 1 2 3 4 5 6 9 8 7 6 5 4 3 2 1 1 2

  8. SXL makes a functional protein in XX zygotes SXL makes a non-functional protein in XY zygotes

  9. L is Late form made During cycle 14 E is Establisment form Made cycle 10 Where is RNA binding domain of SXL protein?

  10. The Sxl protein in different Diptera species. Penalva L O F , and Sánchez L Microbiol. Mol. Biol. Rev. 2003;67:343-359

  11. The Sxl protein in different Diptera species. Penalva L O F , and Sánchez L Microbiol. Mol. Biol. Rev. 2003;67:343-359

  12. The Sxl protein in different Diptera species. Penalva L O F , and Sánchez L Microbiol. Mol. Biol. Rev. 2003;67:343-359

  13. SXL makes a functional protein in XX zygotes SXL makes a non-functional protein in XYzygotes Event is initiated by Exons E1+ 4 No early form of protein, thus no SXL 1/2/4 form SXL 1/2/4 form

  14. Zygotic transcription starts Cycle10-14 Cycle14+

  15. ~Cycle 9 Cycle10-14 Zygotic transcription starts Cycle14-and up

  16. KEY SLIDE…Know it. Zygotic-X chromosome factors:XSEs 2x 1x Gro-maternal Dpn-Zygotic 2x difference Maternal gro Down regulated by Zygotic XSE proteins Zygotic deadpan Represses SxlPe with Maternal gro Development Development

  17. Chapter 19The Genetic Control of Animal Development

  18. Sex Determination in Drosophila and C. elegans • The sex determination signal in both animals is the ratio of X chromosomes to autosomes. If the ratio is 1.0 or greater, the animal is a female; if the ratio is 0.5 or less, the animal is a male.CLASSIC Definition • But wrong • In Drosophila, the key genes in sex determination encode proteins that regulate RNA processing.

  19. Sex Determination in Drosophila • Components of the sex-determination pathway include • A system to ascertain the X:A ratio , • A system to covert this CONCEPTUAL ratio into a developmental signal, and • A system to respond to this signal by producing either male or female structures.

  20. NOT CORRECT..from book Ascertaining the X:A Ratio • The system that ascertains the X:A ratio involves interactions between maternally synthesized proteins in the egg cytoplasm and embryonically synthesized proteins encoded by several X-linked genes. • The X-linked gene products are called numerator elements and are twice as abundant in XX embryos as in XY embryos. • The autosomal gene products are called denominator elements and antagonize the products of the numerator elements- NONE FOUND.

  21. NOT CORRECT..from book

  22. The Sex-lethal (Sxl) Gene • Sxl is the master regular of the sex determination pathway in Drosophila. • The X:A ratio is converted into a molecular signal that controls the expression of the X-linked Sxl gene.

  23. Function of SXL • SXL regulates splicing of its own transcript to maintain SXL protein expression in XX embryos. • SXL also regulates splicing of the transformer (tra) gene.

  24. Differentiating in Response to the Signal • TRA, along with TRA2, regulate splicing of doublesex (dsx) and fruitless (fru). • In XX embryos, where TRA is present, dsx transcripts are processed to encode a DSX protein that represses the genes for male development. • In XY embryos, where TRA is absent, dsx transcripts are processed to encode a DSX protein that represses the genes for female development.

  25. Loss-of-Function Mutations in Sex-Determination Genes in Drosophila • Mutations in Sxl prevent SXL protein from being made in males; homozygous mutants would develop into males but die as embryos. • Mutations in transformer and transformer2 cause both XX and XY animals to develop into males. • Mutations in dsx cause both XX and XY embryos to develop into intersexes.

  26. Key Points • In Drosophila the pathway that controls sexual differentiation involves some genes that ascertain the X:A ratio, some that convert this ratio into a developmental signal, and others that respond to the signal by producing either male or female structures. • The Sex-lethal (Sxl) gene plays a key role in Drosophila sexual development by regulating the splicing of its own transcript and that of another gene (tra).