Post‐translational processing of the amino terminus affects actin function

Emma S. HENNESSEY, Douglas R. DRUMMOND, John C. SPARROW

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

We have studied the importance of N‐terminal processing for normal actin function using the Drosophila Act88F actin gene transcribed and translated in vitro. Despite having different charges as determined by two‐dimensional (2D) gel electrophoresis, Act88F expressed in vivo and in vitro in rabbit reticulocyte lysate bind to DNase I with equal affinity and are able to copolymerise with bulk rabbit actin equally well. Using peptide mapping and thin‐layer electrophoresis we have shown that bestatin ([3‐amino‐2‐hydroxy‐4‐phenyl‐butanoyl]‐l‐leucine), an inhibitor of aminopeptidases, can inhibit actin N‐terminal processing in rabbit reticulocyte lysate. Although processed and unprocessed actins translated in vitro are able to bind to DNase I equally well, unprocessed actins are less able to copolymerise with bulk actins. This effect is more pronounced when bulk rabbit actin is used but is still seen with bulk Lethocerus actin. Also, the unprocessed actins reduce the polymerisation of the processed actin translated in vitro with the bulk rabbit actin. This suggests that individual actins do interact, even in non‐polymerising conditions. The reduced ability of unprocessed actin to polymerise shows that correct post‐translational modification of the N terminus is required for normal actin function.

Original languageEnglish
Pages (from-to)345-352
Number of pages8
JournalEuropean Journal of Biochemistry
Volume197
Issue number2
DOIs
Publication statusPublished - Jan 1 1991
Externally publishedYes

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Actins
Processing
Rabbits
Deoxyribonuclease I
Reticulocytes
Electrophoresis
Aminopeptidases
Peptide Mapping
Electrophoresis, Gel, Two-Dimensional
Post Translational Protein Processing
Polymerization
Drosophila
Genes
Gels

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Post‐translational processing of the amino terminus affects actin function. / HENNESSEY, Emma S.; DRUMMOND, Douglas R.; SPARROW, John C.

In: European Journal of Biochemistry, Vol. 197, No. 2, 01.01.1991, p. 345-352.

Research output: Contribution to journalArticle

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