Expression of P-glycoprotein gene in marine sponges. Identification and characterization of the 125 kDa drug-binding glycoprotein.
In the present paper it is shown that the marine sponges Geodia cydonium and Verongia aerophoba contain the gene coding for P-glycoprotein P170, also known as a multidrug-resistance gene. Western blot studies revealed that polyclonal antibodies raised against hamster P170 cross-react with the sponge polypeptide of Mr 125,000. After endoglycosidase F treatment, the sponge P125 is converted to a polypeptide of Mr 105,000. Northern blot studies, using the human P170 cDNA probe, revealed a size of 4.2 kb for the sponge P125 transcript. The level of this transcript does not change in response to incubation with the aggregation factor. Confocal laser scanning microscopy showed that P125 is a cell membrane bound protein. In addition, sponge membrane vesicles possess a potential to bind in vitro 2-acetylamino-fluorene, vincristine and daunomycin. This process is Verapamil-sensitive, a characteristic known also for the mammalian vesicle associated P170. The data reported demonstrate that the classical multidrug resistance mechanism, described in drug-resistant tumor cell lines, functions also in sponges and may explain the relative resistance of these animals to pollution.
Kurelec
B
B
Center for Marine Research Zagreb, Ruder Bosković Institute, Zagreb, Croatia.
Krca
S
S
Pivcevic
B
B
Ugarković
D
D
Bachmann
M
M
Imsiecke
G
G
Müller
W E
WE
D015073:2-Acetylaminofluorene / Q000378:metabolism; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification*; D003630:Daunorubicin / Q000378:metabolism; D004351:Drug Resistance; D015870:Gene Expression; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000302:isolation & purification; D011161:Porifera / Q000378:metabolism*; D014700:Verapamil / Q000494:pharmacology; D014750:Vincristine / Q000378:metabolism
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D014750:Vincristine; D015073:2-Acetylaminofluorene; D014700:Verapamil; D003630:Daunorubicin