Molecular cloning, sequencing, and brain expression of the presenilin 1 gene in Microcebus murinus.
The cDNA encoding the Microcebus murinus presenilin 1 protein (PSI) was cloned by RT-PCR from a brain cDNA library using various combinations of oligonucleotide primers designed on the basis of the human nucleotide sequence. Analysis of five clones isolated from two positive combinations revealed that the deduced open reading frame encodes two protein isoforms of 467 and 463 amino acid residues. The shorter isoform lacked the four residues VRSQ in the N-terminal region and like the 467 amino acid isoform presented 22 substitutions with its human homologue. The 12 bp nucleotide deletion evidenced in the cDNA encoding the shorter isoform is consistent with the use of an alternative 5' splice donor site identified at the end of the human exon 3. The immunohistochemistry performed with a specific polyclonal antiserum raised against a synthetic peptide located in the human large hydrophilic loop of PS1 revealed that the protein is widely distributed independently of age or of pathology in the microcebe brain. PS1 is found predominantly in neurons of the different cortical layers and hippocampus but also in subcortical structures. The PS1 labelling appeared as thin granulations scattered throughout the cytoplasm of numerous neurons and sometimes in neurites.
Calenda
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A
INSERM U249, CNRS UPR9008, Institut de Biologie, Montpellier, France. calenda@xerxes.crbm.cnrs-mop.fr
Mestre-Francés
N
N
Czech
C
C
Pradier
L
L
Petter
A
A
Bons
N
N
Bellis
M
M
D000375:Aging; D000595:Amino Acid Sequence; D000686:Amyloidosis / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D016565:Cheirogaleidae; D003001:Cloning, Molecular; D017931:DNA Primers; D006801:Humans; D007150:Immunohistochemistry; D007900:Lemur; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D016501:Neurites / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D016366:Open Reading Frames; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D053764:Presenilin-1; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D008565:Membrane Proteins; C508853:PSEN1 protein, human; D053764:Presenilin-1; D011994:Recombinant Proteins