PMID- 10610717
OWN - NLM
STAT- MEDLINE
DCOM- 20000209
LR  - 20120327
IS  - 0888-7543 (Print)
IS  - 0888-7543 (Linking)
VI  - 62
IP  - 2
DP  - 1999 Dec 1
TI  - Molecular cloning and characterization of a human cDNA and gene encoding a novel 
      acid ceramidase-like protein.
PG  - 232-41
AB  - Computer-assisted database analysis of sequences homologous to human acid
      ceramidase (ASAH) revealed a 1233-bp cDNA (previously designated cPj-LTR) whose
      266-amino-acid open reading frame had approximately 36% identity with the ASAH
      polypeptide. Based on this high degree of homology, we undertook further
      molecular characterization of cPj-LTR and now report the full-length cDNA
      sequence, complete gene structure (renamed human ASAHL since it is a human acid
      ceramidase-like sequence), chromosomal location, primer extension and promoter
      analysis, and transient expression results. The full-length human ASAHL cDNA was 
      1825 bp and contained an open-reading frame encoding a 359-amino-acid polypeptide
      that was 33% identical and 69% similar to the ASAH polypeptide over its entire
      length. Numerous short regions of complete identity were observed between these
      two sequences and two sequences obtained from the Caenorhabditis elegans genome
      database. The 30-kb human ASAHL genomic sequence contained 11 exons, which ranged
      in size from 26 to 671 bp, and 10 introns, which ranged from 150 bp to 6.4 kb.
      The gene was localized to the chromosomal region 4q21.1 by fluorescence in situ
      hybridization analysis. Northern blotting experiments revealed a major 2.0-kb
      ASAHL transcript that was expressed at high levels in the liver and kidney, but
      at relatively low levels in other tissues such as the lung, heart, and brain.
      Sequence analysis of the 5'-flanking region of the human ASAHL gene revealed a
      putative promoter region that lacked a TATA box and was GC rich, typical features
      of a housekeeping gene promoter, as well as several tissue-specific and/or
      hormone-induced transcription regulatory sites. 5'-Deletion analysis localized
      the promoter activity to a 1. 1-kb fragment within this region. A major
      transcription start site also was located 72 bp upstream from the ATG translation
      initiation site by primer extension analysis. Expression analysis of a green
      fluorescence protein/ASAHL fusion protein in COS-1 cells revealed a punctate,
      perinuclear distribution, although no acid ceramidase activity was detected in
      the transfected cells using a fluorescence-based in vitro assay system.
CI  - Copyright 1999 Academic Press.
FAU - Hong, S B
AU  - Hong SB
AD  - Department of Human Genetics, Mount Sinai School of Medicine, New York 10029,
      USA.
FAU - Li, C M
AU  - Li CM
FAU - Rhee, H J
AU  - Rhee HJ
FAU - Park, J H
AU  - Park JH
FAU - He, X
AU  - He X
FAU - Levy, B
AU  - Levy B
FAU - Yoo, O J
AU  - Yoo OJ
FAU - Schuchman, E H
AU  - Schuchman EH
LA  - eng
GR  - DK 54830/DK/NIDDK NIH HHS/United States
GR  - HD 28607/HD/NICHD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Genomics
JT  - Genomics
JID - 8800135
RN  - 0 (5' Untranslated Regions)
RN  - 0 (DNA, Complementary)
RN  - 0 (Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 63231-63-0 (RNA)
RN  - EC 3.2.1.47 (Galactosylgalactosylglucosylceramidase)
RN  - EC 3.5.- (Amidohydrolases)
RN  - EC 3.5.1.- (NAAA protein, human)
SB  - IM
MH  - 5' Untranslated Regions/chemistry
MH  - Amidohydrolases/biosynthesis/chemistry/*genetics
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - COS Cells
MH  - Chromosomes, Human, Pair 4/genetics
MH  - Cloning, Molecular
MH  - DNA, Complementary/*chemistry/genetics/isolation & purification
MH  - Galactosylgalactosylglucosylceramidase/*chemistry
MH  - Humans
MH  - Mice
MH  - Molecular Sequence Data
MH  - Organ Specificity/genetics
MH  - Protein Biosynthesis
MH  - Proteins/chemistry/*genetics
MH  - RNA/biosynthesis
MH  - Recombinant Fusion Proteins/biosynthesis
MH  - *Sequence Homology, Amino Acid
MH  - Transcription, Genetic/genetics
MH  - Transfection
EDAT- 1999/12/28 00:00
MHDA- 1999/12/28 00:01
CRDT- 1999/12/28 00:00
PHST- 1999/12/28 00:00 [pubmed]
PHST- 1999/12/28 00:01 [medline]
PHST- 1999/12/28 00:00 [entrez]
AID - 10.1006/geno.1999.5953 [doi]
AID - S0888-7543(99)95953-0 [pii]
PST - ppublish
SO  - Genomics. 1999 Dec 1;62(2):232-41. doi: 10.1006/geno.1999.5953.