Changes in cysteine protease activity and localization during midgut metamorphosis in the crucifer root maggot (Delia radicum).
We show that differential localization and/or activation of two cysteine protease activities occur at the onset of dipteran midgut metamorphosis. A 26 kDa cysteine protease activity was associated specifically with midgut tissues of late third instar larvae. Starvation of mid third instar larvae simulated the onset of prepupation and resulted in loss of the 26 kDa protease activity. A cDNA clone encoding a cysteine protease, termed DrCP1, was isolated and shown to be highly similar to those from Sarcophaga peregrina and Drosophila melanogaster (DmCP1). DrCP1 mRNA was present in all developmental stages including eggs, larvae, pupae and adults, but was highly induced at the onset of the larval-pupal transition and thereafter. The DrCP1 protein is localized to the exterior of the midgut tissues during the onset of the prepupal transition period, possibly in response to ecdysone. Analysis of transcription factor binding sites associated with the DmCP1 promoter indicated that elements exist that allow for both ecdysone-mediated as well as tissue-specific regulation. Based upon these and other studies we propose: (1) that the expression, activity and localization of the DrCP1-like cysteine proteases are highly regulated throughout development; and, (2) that cysteine protease activities are involved in aspects of tissue reconstruction at the onset of and during metamorphosis.
Hegedus
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Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon SK, Canada S7N 0X2.
O'Grady
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Chamankhah
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Baldwin
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Gleddie
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Braun
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Erlandson
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D004064:Digestive System / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015723:Gene Library; D006801:Humans; D007814:Larva; D008322:Mammals; D008675:Metamorphosis, Biological; D008969:Molecular Sequence Data; D009117:Muscidae / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D010944:Plants / Q000201:enzymology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D003546:Cysteine Endopeptidases