Genomic organization and molecular characterization of a gene encoding HsPXF, a human peroxisomal farnesylated protein.
A protein modification essential for the cellular sorting of many biologically relevant proteins is the covalent attachment of prenyl lipids by specific transferases. Isoprenylation is known to render protein domains hydrophobic, thereby facilitating the interaction with lipid bilayers and/or membrane proteins. The target for the modification with farnesyl groups is the COOH-terminal sequence CaaX. Among the variety of farnesylated proteins the only one reported so far to be located to peroxisomes is the 37-kDa peroxisomal farnesylated hamster protein PxF. Recently we published data on the cDNA of the human gene HK33 (A. Braun et al., 1994, Gene 146: 291-295), which was revealed to be the human ortholog of PxF and was consequently renamed HsPXF. The genomic structure, molecular characterization, and evolutionary conservation of HsPXF are described herein. The exact location of the gene was defined as chromosome 1q22. The gene spans a region of approximately 9 kb, containing eight exons and seven introns. The 5' upstream region showed two potential Sp1-binding sites and an Alu repetitive sequence. Luciferase reporter activating capacity confirmed the presumed promoter activity of this region. On the transcriptional level, we detected four splice variants originating either from exon skipping or from alternative splicing events. For the HsPXF protein, a carboxyterminal farnesylation at cysteine residues was demonstrated. Through the use of HsPXF-specific antibodies, the protein was shown to be attached to the outer surface of peroxisomes. This localization together with the similarity to a peroxisomal assembly protein from Saccharomyces cerevisiae suggests HsPXF is involved in the process of peroxisomal biogenesis or assembly.
Kammerer
S
S
Institute of Anthropology and Human Genetics, University of Munich, Germany.
Arnold
N
N
Gutensohn
W
W
Mewes
H W
HW
Kunau
W H
WH
Höfler
G
G
Roscher
A A
AA
Braun
A
A
D000595:Amino Acid Sequence; D000818:Animals; D002874:Chromosome Mapping; D002878:Chromosomes, Human, Pair 1; D006224:Cricetinae; D018076:DNA, Complementary; D019143:Evolution, Molecular; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008830:Microbodies / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012326:RNA Splicing; D013347:Subcellular Fractions / Q000378:metabolism
D018076:DNA, Complementary; D008565:Membrane Proteins; C087343:PEX19 protein, human