Expression cloning of LDLB, a gene essential for normal Golgi function and assembly of the ldlCp complex

J. E. Chatterton, D. Hirsch, J. J. Schwartz, P. E. Bickel, R. D. Rosenberg, H. F. Lodish, M. Krieger

Research output: Contribution to journalArticle

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Abstract

The Chinese hamster ovary (CHO) cell mutants ldlC and ldlB, which exhibit almost identical phenotypes, define two genes required for multiple steps in the normal medial and trans Golgi-associated processing of glycoconjugates. The LDLC gene encodes IdlCp, an ≃80-kDa protein, which in wild-type, but not ldlB, cells associates reversibly with the cytoplasmic surface of the Golgi apparatus. Here, we have used a retrovirus-based expression cloning system to clone a murine cDNA, LDLB, that corrects the pleiotropic mutant phenotypes of ldlB cells. The corresponding mRNA was not detected in ldlB mutants. LDLB encodes an ≃110-kDa protein, ldlBp, which lacks homology to known proteins and contains no common structural motifs. Database searches identified short segments of homology to sequences from Drosophila melanogaster, Arabidopsis thaliana, and Caenorhabditis elegans, and the essentially full-length homologous human sequence (82% identity); however, as was the case for ldlCp, no homologue was identified in Saccharomyces cerevisiae. We have found that in wild-type cell cytosols, ldlCp is a component of an ≃950-kDa 'ldlCp complex,' which is smaller, ≃700 kDa, in ldlB cytosols. Normal assembly of this complex is ldlBp-dependent and may be required for Golgi association of ldlCp and for the normal activities of multiple luminal Golgi processes.

Original languageEnglish (US)
Pages (from-to)915-920
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number3
DOIs
StatePublished - Feb 2 1999

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Essential Genes
Organism Cloning
Sequence Homology
Cytosol
Phenotype
Proteins
Glycoconjugates
Caenorhabditis elegans
Golgi Apparatus
Retroviridae
Cricetulus
Drosophila melanogaster
Arabidopsis
Genes
Saccharomyces cerevisiae
Ovary
Complementary DNA
Clone Cells
Databases
Messenger RNA

ASJC Scopus subject areas

  • Genetics
  • General

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Expression cloning of LDLB, a gene essential for normal Golgi function and assembly of the ldlCp complex. / Chatterton, J. E.; Hirsch, D.; Schwartz, J. J.; Bickel, P. E.; Rosenberg, R. D.; Lodish, H. F.; Krieger, M.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 3, 02.02.1999, p. 915-920.

Research output: Contribution to journalArticle

Chatterton, J. E. ; Hirsch, D. ; Schwartz, J. J. ; Bickel, P. E. ; Rosenberg, R. D. ; Lodish, H. F. ; Krieger, M. / Expression cloning of LDLB, a gene essential for normal Golgi function and assembly of the ldlCp complex. In: Proceedings of the National Academy of Sciences of the United States of America. 1999 ; Vol. 96, No. 3. pp. 915-920.
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