Protein patterns in the oat coleoptile as influenced by auxin and by protein turnover

Abstract

Synthesis of growth-limiting proteins (GLP) is required for continued auxin-induced elongation of oat (Avena sativa L.) coleoptiles. In order to determine whether GLP synthesis is dependent or independent of auxin, a double-labeling ratio technique, coupled with disc-gel electrophoresis, has been used to assess the effect of auxin on the pattern of protein synthesis. Sections were peeled to enhance amino-acid uptake; proteins were labeled with [¹⁴C]- or [³H] leucine in the presence or absence of indole-3-acetic acid for 40 min to 6 h, and were separated into soluble, membrane-associated, and wall-associated fractions. Regardless of the conditions used, or the protein fraction examined, no changes in response to auxin were detected in the pattern of protein synthesis. In order to escape detection by this technique an auxin-induced protein would have to comprise less than 0.75% of the total newly synthesized protein. Thus the synthesis of GLP appears to be independent of auxin. The same technique has been used to follow protein turnover. During the chase, proteins are initially degraded at an average rate of 8% h⁻¹, and some protein bands showed as much as 14% h⁻¹ degradation. No protein was detected which had a turnover rate as rapid as the GLP.