J Cell Biol. 1993 Sep;122(6):1301-10. doi: 10.1083/jcb.122.6.1301.

Chaperonin-mediated folding of vertebrate actin-related protein and gamma-tubulin.

The Journal of cell biology

R Melki, I E Vainberg, R L Chow, N J Cowan

PMID: 8104191 PMCID: PMC2119862 DOI: 10.1083/jcb.122.6.1301
Free PMC Article

Abstract

The folding of actin and tubulin is mediated via interaction with a heteromeric toroidal complex (cytoplasmic chaperonin) that hydrolyzes ATP as part of the reaction whereby native proteins are ultimately released. Vertebrate actin-related protein (actin-RPV) (also termed centractin) and gamma-tubulin are two proteins that are distantly related to actin and tubulin, respectively: gamma-tubulin is exclusively located at the centrosome, while actin-RPV is conspicuously abundant at the same site. Here we show that actin-RPV and gamma-tubulin are both folded via interaction with the same chaperonin that mediates the folding of beta-actin and alpha- and beta-tubulin. In each case, the unfolded polypeptide forms a binary complex with cytoplasmic chaperonin and is released as a soluble, monomeric protein in the presence of Mg-ATP and the presence or absence of Mg-GTP. In contrast to alpha- and beta-tubulin, the folding of gamma-tubulin does not require the presence of cofactors in addition to chaperonin itself. Monomeric actin-RPV produced in in vitro folding reactions cocycles efficiently with native brain actin, while in vitro folded gamma-tubulin binds to polymerized microtubules in a manner consistent with interaction with microtubule ends. Both monomeric actin-RPV and gamma-tubulin bind to columns of immobilized nucleotide: monomeric actin-RPV has no marked preference for ATP or GTP, while gamma-tubulin shows some preference for GTP binding. We show that actin-RPV and gamma-tubulin compete with one another, and with beta-actin or alpha-tubulin, for binary complex formation with cytoplasmic chaperonin.

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Substances

• Actins
• Proteins
• Tubulin
• centractin
• Guanosine Triphosphate
• Adenosine Triphosphate
• Chaperonins

MeSH terms

• Actins / chemistry
• Actins / metabolism
• Adenosine Triphosphate / metabolism
• Animals
• Base Sequence
• Binding, Competitive
• Cattle
• Chaperonins
• Guanosine Triphosphate / metabolism
• Male
• Mice
• Molecular Sequence Data
• Peptide Biosynthesis
• Protein Binding
• Protein Conformation
• Proteins / metabolism
• Proteins / physiology
• Rabbits
• Reticulocytes / chemistry
• Reticulocytes / metabolism
• Tubulin / chemistry
• Tubulin / metabolism

Publication Types

• Journal Article
• Research Support, Non-U.S. Gov't
• Research Support, U.S. Gov't, P.H.S.

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