Evolution and mechanism of spectral tuning of blue-absorbing visual pigments in butterflies.
The eyes of flower-visiting butterflies are often spectrally highly complex with multiple opsin genes generated by gene duplication, providing an interesting system for a comparative study of color vision. The Small White butterfly, Pieris rapae, has duplicated blue opsins, PrB and PrV, which are expressed in the blue (λ(max) = 453 nm) and violet receptors (λ(max) = 425 nm), respectively. To reveal accurate absorption profiles and the molecular basis of the spectral tuning of these visual pigments, we successfully modified our honeybee opsin expression system based on HEK293s cells, and expressed PrB and PrV, the first lepidopteran opsins ever expressed in cultured cells. We reconstituted the expressed visual pigments in vitro, and analysed them spectroscopically. Both reconstituted visual pigments had two photointerconvertible states, rhodopsin and metarhodopsin, with absorption peak wavelengths 450 nm and 485 nm for PrB and 420 nm and 482 nm for PrV. We furthermore introduced site-directed mutations to the opsins and found that two amino acid substitutions, at positions 116 and 177, were crucial for the spectral tuning. This tuning mechanism appears to be specific for invertebrates and is partially shared by other pierid and lycaenid butterfly species.
Laboratory of Neuroethology, Sokendai-Hayama, Hayama, Japan.
Motohiro
M
Wakakuwa
Akihisa
A
Terakita
Mitsumasa
M
Koyanagi
Doekele G
DG
Stavenga
Yoshinori
Y
Shichida
Kentaro
K
Arikawa
D000595:Amino Acid Sequence; D000818:Animals; D002080:Butterflies / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D019143:Evolution, Molecular; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D015151:Immunoblotting; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012168:Retinal Pigments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017299:Rod Opsins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry / Q000379:methods
D011994:Recombinant Proteins; D012168:Retinal Pigments; D017299:Rod Opsins; C095853:short-wavelength opsin