Cloning of the cDNA and mRNA expression of CLRP, a complex leucine repeat protein of the Golgi apparatus expressed by specific neurons of the rat brain.
We report the molecular cloning of one novel cDNA isolated from the rat brain. We have named the putative protein CLRP, for complex leucine-repeat protein. The predicted CLRP amino acid sequence shares homology in the amino acid composition with the Galactose, N-Acetylglucosamine, and Sialic acid transporters, and shows 91% identity with the sequence of one human chromosome 5 BAC clone. Expression of the CLRP cDNA tagged with GFP in COS-7 cells was found in cell organelles that resemble the Golgi apparatus of the cytoplasm. In Northern blot, the CLRP probe labels a single band of 2.4 kb in the brain, kidney, lung, testis, and prostate. In the brain, CLRP mRNA is expressed by limited sets of neurons, such as the pyramidal cells of the cortex, the Purkinje cells of the cerebellum, and the motoneurons of the brainstem. In the brain, the CLRP mRNA is expressed at embryonic day 15; levels of expression are maintained until postnatal day 10 and decrease in adults. The results suggest that CLRP codes a novel member of the nucleotide-sugar family of proteins of the Golgi apparatus.
Department of Cell Biology and Genetics, Alcalá de Henares University, 28871, Madrid, Spain. julio.pmarquez@uah.es
Julio
J
Pérez-Márquez
Begoña
B
Reguillo
Ricardo
R
Paniagua
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000166:cytology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006056:Golgi Apparatus / Q000737:chemistry / Q000502:physiology*; D007930:Leucine / Q000235:genetics; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics*; D009474:Neurons / Q000737:chemistry / Q000502:physiology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D012091:Repetitive Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C465421:Slc35a4 protein, rat; D007930:Leucine