ISSN:
1432-2048
Keywords:
Auxin-binding protein
;
Chaperone
;
Endoplasmic reticulum
;
Plasma membrane
;
Protein synthesis/turnover
;
Zea
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Electrophysiological experiments have indicated that a fraction of the major auxin-binding protein (ABP1) of maize (Zea mays L.) might be a receptor on the outer surface of the plasma membrane. The predominant location of ABP1 is in the lumen of the endoplasmic reticulum (ER), in accord with its C-terminal KDEL retention signal. Little is known about the biology of the protein in vivo or the rate at which it might pass to the cell surface. We have examined the turnover of ABP1 by in vivo labelling of maize coleoptile sections. After different chase times, ABP1 was immunoprecipitated from detergentsolubilised membrane preparations. Two polypeptides coprecipitated with ABP1. Neither was recognised by any ABP1 antibodies nor by monoclonals to ER retention sequences. The possible significance of these coprecipitating polypeptides is discussed. In addition, we have used a monoclonal antibody to precipitate HDEL proteins from the same membrane preparations. Two dimensional electrophoresis and N-terminal sequencing showed that the major HDEL protein precipitated was a member of the heat-shock-protein 70 family, a homologue of BiP (immunoglobulin-binding protein). We have investigated the turnover of this BiP homologue for comparison with ABP1 and found that both had extended lifetimes, with half-lives greater than 24 h. Use of cordycepin to inhibit transcription indicated that ABP1 mRNA was also longlived. Synthesis of ABP1 was strongly reduced by heat stress, was reduced a little in response to dithiothreitol and was not markedly changed by tunicamycin. In contrast, BiP synthesis increased markedly in response to tunicamycin and dithiothreitol and increased a little after heat stress. Neither auxin nor other plant growth regulators altered ABP1 synthesis or turnover. Auxin binding is thought to induce a conformational change in ABP1 which might be associated with its escape from the ER. Incubation of coleoptile sections in auxin did not alter the balance of ABP1 between ER-enriched and plasmamembrane-enriched fractions over prolonged chase times. We discuss the turnover characteristics of ABP1 in terms of its known cellular distribution and of its function.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00196668
