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Related Concept Videos

Mutations01:39

Mutations

Overview
Mutations01:39

Mutations

Overview
Tail-anchoring of Proteins in the ER Membrane01:45

Tail-anchoring of Proteins in the ER Membrane

Tail-anchored, or TA, proteins are estimated to make up to 3-5% of membrane proteins found in the eukaryotic cell. Such proteins have a single transmembrane domain located approximately 30 amino acid residues upstream from the C-terminal end. As a result, the signal recognition particle (SRP) cannot guide a TA protein to the ER membrane for cotranslational insertion. Hence, they are integrated into the ER membrane post-translationally using their C-terminal end as the anchor. TA proteins...
Mutations01:35

Mutations

Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations can be characterized in a number of different ways: whether and how they alter the amino acid sequence of the protein, whether they occur over a small or large area of DNA, and whether they occur in somatic cells or germline cells.
Chromosomal Alterations Are Large-Scale Mutations
While point mutations are changes in a single nucleotide in...
Spontaneous and Induced Mutations01:30

Spontaneous and Induced Mutations

Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).
Point and Frameshift Mutations01:30

Point and Frameshift Mutations

Point mutations are genetic alterations involving the change of a single nucleotide base pair in DNA. Depending on how the alteration affects protein synthesis, they can lead to various consequences.Point mutations fall into the following types:Silent mutations occur when a nucleotide change does not alter the amino acid sequence due to the redundancy of the genetic code. For instance, changing ACC to ACA still encodes threonine, leaving the protein function unaffected. This occurs because...

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Related Experiment Video

Updated: Jul 19, 2026

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells
15:28

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Published on: October 1, 2010

Characterization of the sec1-1 and sec1-11 mutations.

M H Brummer1, K J Kivinen, J Jäntti

  • 1VTT Biotechnology, PO Box 1500, FIN-02044 VTT Espoo, Finland.

Yeast (Chichester, England)
|December 19, 2001
PubMed
Summary

Temperature-sensitive mutations in Sec1 proteins (sec1-1 and sec1-11) alter protein folding and interaction with SNARE complex partners. These mutations lead to protein instability and loss of function at higher temperatures.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Protein Biochemistry

Background:

  • Sec1 proteins regulate SNARE complex formation, crucial for vesicle trafficking.
  • Understanding Sec1 protein function requires characterizing mutations affecting its activity.

Purpose of the Study:

  • To identify the molecular basis of temperature-sensitive mutations sec1-1 and sec1-11.
  • To elucidate the impact of these mutations on Sec1 protein structure, stability, and function.

Main Methods:

  • Site-directed mutagenesis to create sec1-1 (G443E) and sec1-11 (R432P) mutations.
  • Protein abundance and stability assays (pulse-chase).
  • Yeast two-hybrid assays to assess protein-protein interactions.

Main Results:

  • Mutations G443E and R432P localize to domain 3b of Sec1p.
  • Mutant Sec1 proteins show reduced abundance and impaired interaction with Mso1p and Sso2p even at permissive temperatures.
  • Mutant proteins are non-functional at 37°C, suggesting slow folding and susceptibility to degradation before proper complex formation.

Conclusions:

  • The sec1-1 and sec1-11 mutations confer temperature sensitivity by affecting Sec1 protein folding and interaction with SNARE partners.
  • These findings provide insights into the structural requirements for Sec1 protein function and regulation of SNARE complex assembly.