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Related Experiment Videos

LDLC encodes a brefeldin A-sensitive, peripheral Golgi protein required for normal Golgi function

S D Podos1, P Reddy, J Ashkenas

  • 1Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

The Journal of Cell Biology
|November 1, 1994
PubMed
Summary

Researchers identified a new Golgi protein, ldlCp, crucial for multiple Golgi functions. Its association with the Golgi depends on the LDLB gene and brefeldin A, highlighting its role in cellular processes.

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Area of Science:

  • Cell Biology
  • Molecular Genetics
  • Glycobiology

Background:

  • Low density lipoprotein (LDL) receptor-deficient Chinese hamster ovary (CHO) cell mutants (ldlB, ldlC) display defects in Golgi-associated processes.
  • Abnormal glycosylation in these mutants leads to reduced LDL receptor stability and activity.

Purpose of the Study:

  • To identify the gene responsible for the ldlC mutant phenotype.
  • To characterize the function and localization of the protein encoded by the identified gene.

Main Methods:

  • cDNA cloning and sequencing of a human gene (LDLC) that corrects ldlC cell defects.
  • Analysis of LDLC mRNA levels in wild-type and mutant cells.
  • Protein sequence prediction and comparison.
  • Immunofluorescence studies to determine protein localization.

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Main Results:

  • A human cDNA (LDLC) was cloned and sequenced, correcting ldlC cell defects but not ldlB.
  • ldlC cells lack detectable LDLC mRNA, identifying LDLC as the defective gene.
  • The predicted protein, ldlCp, is novel, lacking known structural motifs.
  • ldlCp is a peripheral Golgi protein, sensitive to brefeldin A.
  • In ldlB cells, ldlCp is not associated with the Golgi, indicating LDLB dependence.

Conclusions:

  • A previously unrecognized protein, ldlCp, is essential for multiple Golgi functions.
  • ldlCp's peripheral association with the Golgi is dependent on the LDLB gene and brefeldin A.
  • This discovery provides new insights into Golgi-associated cellular processes and genetic regulation.