Thyrotropin stimulation of lysosomal tyrosine transport in rat FRTL-5 thyroid cells.
Tyrosine countertransport was used to demonstrate the hormonal stimulation of neutral amino acid transport across the lysosomal membrane of FRTL-5 cells. Cells grown with thyrotropin (1 X 10(-10) M) had 7-fold (+/- S.E.) higher tyrosine countertransport activity in their lysosomes than cells grown without thyrotropin. Thyrotropin also stimulated the uptake into tyrosine-loaded lysosomes of other neutral amino acids recognized by the tyrosine carrier, namely, phenylalanine (3-fold) and leucine (6-fold). In contrast lysosomal cystine countertransport was not affected by thyrotropin. Addition of thyrotropin to cells grown without thyrotropin showed that the stimulation of tyrosine counter-transport (a) required at least 48 h to reach the level of the thyrotropin-supplemented cells, (b) depended upon protein synthesis, since cycloheximide (20 microM) was inhibitory, and (c) depended upon RNA synthesis, since actinomycin D (1 nM) was inhibitory. Cells grown without thyrotropin but with dibutyryl cyclic AMP (1 mM) or cholera toxin (1 nM) exhibited enhanced lysosomal countertransport of tyrosine, suggesting that cyclic AMP may act as a messenger. This represents the first demonstration of hormonal responsiveness in a lysosomal transport system and may reflect the importance of salvage and reutilization of lysosomal degradation products for the thyroid epithelial cell.
Harper
G S
GS
Section on Human Biochemical Genetics, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
Kohn
L D
LD
Bernardini
I
I
Bernar
J
J
Tietze
F
F
Andersson
H C
HC
Gahl
W A
WA
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D003994:Bucladesine / Q000494:pharmacology; D002460:Cell Line; D002772:Cholera Toxin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003553:Cystine / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism; D051381:Rats; D013961:Thyroid Gland; D013972:Thyrotropin / Q000494:pharmacology*; D014443:Tyrosine / Q000378:metabolism*
D003609:Dactinomycin; D014443:Tyrosine; D010649:Phenylalanine; D003553:Cystine; D003994:Bucladesine; D013972:Thyrotropin; D002772:Cholera Toxin; D003513:Cycloheximide; D000242:Cyclic AMP; D007930:Leucine