Functional and physiological consequences of genetic variation at phosphoglucose isomerase: Heat shock protein expression is related to enzyme genotype in a montane beetle

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Abstract
Allele frequency variation at the phosphoglucose isomerase (PGI) locus in Californian populations of the beetle Chrysomela aeneicollis suggests that PGI may be undergoing natural selection. We quantified (i) apparent Michaelis-Menten constant (K(m)) of fructose 6-phosphate at different temperatures and (ii) thermal stability for three common PGI genotypes (1-1, 1-4, and 4-4). We also measured air temperature (T(a)) and beetle body temperature (T(b)) in three montane drainages in the Sierra Nevada, California. Finally, we measured 70-kDa heat shock protein (Hsp70) expression in field-collected and laboratory-acclimated beetles. We found that PGI allele 1 predominated in the northernmost drainage, Rock Creek (RC), which was also significantly cooler than the southernmost drainage, Big Pine Creek (BPC), where PGI allele 4 predominated. Allele frequencies and air temperatures were intermediate in the middle drainage, Bishop Creek (BC). Differences among genotypes in K(m) (1-1 > 1-4 > 4-4) and thermal stability (4-4 > 1-4 > 1-1) followed a pattern consistent with temperature adaptation. In nature, T(b) was closely related to T(a). Hsp70 expression in adult beetles decreased with elevation and differed among drainages (BPC > BC > RC). After laboratory acclimation (8 days, 20 degrees C day, 4 degrees C night) and heat shock (4 h, 28-36 degrees C), Hsp70 expression was greater for RC than BPC beetles. In RC, field-collected beetles homozygous for PGI 1-1 had higher Hsp70 levels than heterozygotes or a 4-4 homozygote. These results reveal functional and physiological differences among PGI genotypes, which suggest that montane populations of this beetle are locally adapted to temperature.