Lecture 24 Protein Translocation, Organellar Genomes

Page 1: Overview

  • Lecture 24 Topics:

    • Protein Translocation

    • Organellar Genomes

Page 2: Learning Goals

  • Understand how nuclearly-encoded proteins reach their destination:

    • Cytoplasmic Proteins

    • Nuclear Proteins

    • Organellar Proteins

    • Exported Proteins and Endomembrane System

  • Identify organelles with their own genomes and the fate of ancestral genomes of mitochondrion and chloroplast.

Page 3: Protein Synthesis Basics

  • Cytosolic proteins synthesized in the cytosol remain there.

  • Other proteins require embedding in membranes or compartments, which begins with protein synthesis in cytoplasm.

  • Nuclearly-encoded proteins cannot pass through the lipid bilayer directly.

Page 4: Membrane Insertion

  • Some proteins are amphipathic, embedded in membranes.

  • Mechanism for crossing plasma membrane for exported proteins is discussed.

  • Most organellar genes are nuclearly encoded; proteins synthesized in cytoplasm must find their way to organelles.

Page 5: Nuclear Protein Transfer

  • Does nuclear transfer require crossing a membrane?

    • Yes, through the nuclear membrane (both inner and outer).

    • Nuclear pores allow diffusion for small molecules and proteins.<br>

Page 6: Classes of Protein Translocation

  • Two classes of translocation:

    • Diffusion and Translocation

      • N-terminal signal sequence helps in localization.

    • Nuclear localization signals guide nuclear proteins via importins.

  • Post-translational and co-translational translocation methods discussed.

Page 7: Nuclear Pores

  • Structure of nuclear pores facilitates import and export of proteins.

  • Large protein complexes and nuclear localization signals are essential for transport.

  • Examples of proteins that function in the nucleus are mentioned.

Page 8: Imported Proteins

  • Fully folded proteins are imported via nuclear import receptors.

  • Similar receptors exist for RNA export from the nucleus.

Page 9: Nuclear Pore Function

  • Receptor-cargo complexes navigate nuclear pore proteins through a gel-like mesh.

Page 10: Recent Pore Structures

  • Structures of nuclear pores discussed with references provided.

Page 11: Protein Transport to Mitochondria and Plastids

  • Post-translational sorting with signal peptides recognized by transport channels.

Page 12: Mitochondrial Protein Import

  • Proteins synthesized in the cytoplasm remain unfolded while being imported into mitochondria.

Page 13: Targeting Proteins to Thylakoids

  • Proteins must be recognized by sequences and cleaved post-insertion in thylakoids.

Page 14: Example - Thylakoid Protein Import

  • Description of how proteins cross membranes through two channels in thylakoids.

Page 15: Endomembrane System Overview

  • The endomembrane system's various components like nucleas, plasma membrane, golgi, and ER.

Page 16: Endomembrane System Continuity

  • The nuclear lumen is contiguous with the rough ER; protein synthesis begins in the cytosol.

Page 17: Signal Sequences and Translation

  • Signal sequences guide the nascent protein to the rough ER and partner with SRPs for transport; translation proceeds simultaneously.

Page 18: SRPs and Translocation

  • SRPs mediate attachment to the ER membrane via receptors and translocon for protein passage.

Page 19: Role of Signal Sequence

  • Signal sequences enable channel opening and are cleaved post-translocation, allowing protein folding.

Page 20: ER Transmembrane Proteins

  • Additional start/stop transfer sequences regulate transmembrane protein integration into membranes.

Page 21: Topologies of Multi-pass Proteins

  • Design of transmembrane proteins is influenced by internal sequences determining orientation.

Page 22: Golgi Budding and Cargo Transport

  • Mechanism of vesicles from ER to Golgi is detailed, emphasizing microtubule facilitation.

Page 23: Sorting in the Endomembrane System

  • Similar destination signal sequences cause sorting in the ER and Golgi.

Page 24: Quiz on Endomembrane System

  • Question posed regarding identifying components of the endomembrane system.

Page 25: Endomembrane Synthesis and Sorting

  • Proteins synthesized in rough ER, processed, modified and sorted in Golgi with recommended resources for learning.

Page 26: Organellar Genomes and Import Mechanism

  • Organelles have reduced genomes; nuclearly-encoded proteins are required for their function.

Page 27: Gene Migration to the Nucleus

  • Most mitochondrial genes migrated to the nuclear genome; implications discussed.

Page 28: Expression Differences in Gene Transfer

  • Differences between prokaryotic and eukaryotic gene expression highlight challenges following transfer.

Page 29: Expression Hurdles

  • Steps required to express transferred bacterial genes within eukaryotic cells outlined.

Page 30: Current Transfer Mechanisms

  • Modern processes of gene transfer into the nucleus and necessary adaptations are discussed.

Page 31: Gene Transfer Theories

  • Various scenarios of gene migration from mitochondrion to nucleus considered.

Page 32: Reasons for Gene Migration

  • Hypotheses explored regarding the motivation for organellar genes transitioning to the nucleus.

Page 33: DNA Sequence Changes

  • Different sequences highlighted for redundancy without functional change.

Page 34: Mutation Impact

  • Discussion on the effects of specific mutations, corroborated with examples.

Page 35: Functional Impact of Amino Acid Changes

  • Assessing consequences of mutation changes within proteins, emphasizing conditions for impact.

Page 36: Knowledge Application

  • Considerations for understanding mutation impacts based on protein function.

Page 37: Case Study - Mutations with Effects

  • White tiger mutation leading to loss of transporter function as specific example.

Page 38: Nonsense Mutations

  • Nonsense mutation results in incomplete protein synthesis, outlining genetic implications.

Page 39: Mutation Questions

  • Queries surrounding the potential dominance of nonsense mutations discussed.

Page 40: Frame-shift Mutations

  • Consequences of insertions and deletions on amino acid sequence presentation.

Page 41: Frameshift Effects Explained

  • Simple explanations on how frameshift mutations can cause widespread effects on protein structure.

Page 42: Mitigation of Insertions/Deletions

  • Potential resiliency of certain insertion/deletion mutations addressed.

Overview of Protein Translocation and Organellar Genomes

  • Learning Goals: Understand how nuclearly-encoded proteins reach their destinations, including cytoplasmic, nuclear, organellar, and exported proteins.

  • Protein Synthesis: Cytosolic proteins remain in the cytosol, while others need membrane embedding. Nuclearly-encoded proteins can't pass through lipid bilayers directly.

  • Nuclear Protein Transfer: Requires crossing the nuclear membrane via nuclear pores, permitting small molecule diffusion.

  • Translocation Classes: Involves diffusion and translocation, with N-terminal signals and nuclear localization signals guiding proteins.

  • Mitochondrial and Plastid Import: Proteins are synthesized in the cytoplasm in an unfolded state before import, needing specific signal peptides.

  • Endomembrane System: Comprises nuclear, plasma membrane, Golgi, and ER; proteins synthesized in rough ER are processed in the Golgi.

  • Gene Migration: Most mitochondrial genes migrated to the nuclear genome, impacting expression in eukaryotic cells and presenting challenges.