Structure of the human perforin gene. A simple gene organization with interesting potential regulatory sequences

Mathias G Lichtenheld, E. R. Podack

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

We have cloned the human perforin (P1) gene and sequenced 6.2-kb genomic DNA, containing 1.4-kb 5'-flanking region, the 5' untranslated region, the complete coding region and the beginning of the 3' untranslated region. The P1 gene including at least 95-bp 3' untranslated region is organized in only three exons: the first exon (97 bp) contains all but four nucleotides of the 5' untranslated region and was determined by primer extension and S1 nuclease mapping. This exon is separated by 1.7 kb from the second exon containing the remaining (4 bp) 5' untranslated region, the leader peptide and the N-terminal region of P1 up to - but not including - the C9 homologous region. The third exon is separated by a 1.2-kb intron and contains the remainder of the molecule, including at least 90 bp of the 3' untranslated region. This simple gene organization differs from that of the more complicated C9 gene. Because of the unusual intron in the 5' untranslated sequence the transcription initiation (cap) site is located almost 1.8 kb upstream of the ATG start signal. The more immediate 5' flanking sequence contains a CCAAT and GC box but lacks other known promoter elements. Instead, we find three different sequence repeats. One of them, a hexanucleotide sequence with the consensus GCCCTG of unknown significance occurs 19 times within a stretch of 240 bp. Further upstream we localized sequences homologous to the following enhancer and promoter elements: c-fos proto-oncogen, IFN-γ and phorbol ester response elements, five cAMP response elements, and three motifs corresponding to general inducer elements. In addition, a sequence conserved in the 5' flanking region of several T cell genes was identified. The 5' flanking regions of P1, CCP1 (granzyme B) and CCP2 (granzyme C) (kindly provided by Dr. Bleackley) contain as only significant homology cAMP response elements. These findings are consistent with a tight control and regulation of P1, which appears to be distinct from that of granzymes.

Original languageEnglish
Pages (from-to)4267-4274
Number of pages8
JournalJournal of Immunology
Volume143
Issue number12
StatePublished - Dec 1 1989

Fingerprint

Perforin
5' Flanking Region
Exons
5' Untranslated Regions
Response Elements
3' Untranslated Regions
Granzymes
Genes
Introns
Conserved Sequence
Transcription Initiation Site
Consensus Sequence
Phorbol Esters
Sequence Homology
Protein Sorting Signals
Carcinogens
Nucleotides
T-Lymphocytes
DNA

ASJC Scopus subject areas

  • Immunology

Cite this

Structure of the human perforin gene. A simple gene organization with interesting potential regulatory sequences. / Lichtenheld, Mathias G; Podack, E. R.

In: Journal of Immunology, Vol. 143, No. 12, 01.12.1989, p. 4267-4274.

Research output: Contribution to journalArticle

@article{c2a11cf45f854527a3e2ff32e2ac2946,
title = "Structure of the human perforin gene. A simple gene organization with interesting potential regulatory sequences",
abstract = "We have cloned the human perforin (P1) gene and sequenced 6.2-kb genomic DNA, containing 1.4-kb 5'-flanking region, the 5' untranslated region, the complete coding region and the beginning of the 3' untranslated region. The P1 gene including at least 95-bp 3' untranslated region is organized in only three exons: the first exon (97 bp) contains all but four nucleotides of the 5' untranslated region and was determined by primer extension and S1 nuclease mapping. This exon is separated by 1.7 kb from the second exon containing the remaining (4 bp) 5' untranslated region, the leader peptide and the N-terminal region of P1 up to - but not including - the C9 homologous region. The third exon is separated by a 1.2-kb intron and contains the remainder of the molecule, including at least 90 bp of the 3' untranslated region. This simple gene organization differs from that of the more complicated C9 gene. Because of the unusual intron in the 5' untranslated sequence the transcription initiation (cap) site is located almost 1.8 kb upstream of the ATG start signal. The more immediate 5' flanking sequence contains a CCAAT and GC box but lacks other known promoter elements. Instead, we find three different sequence repeats. One of them, a hexanucleotide sequence with the consensus GCCCTG of unknown significance occurs 19 times within a stretch of 240 bp. Further upstream we localized sequences homologous to the following enhancer and promoter elements: c-fos proto-oncogen, IFN-γ and phorbol ester response elements, five cAMP response elements, and three motifs corresponding to general inducer elements. In addition, a sequence conserved in the 5' flanking region of several T cell genes was identified. The 5' flanking regions of P1, CCP1 (granzyme B) and CCP2 (granzyme C) (kindly provided by Dr. Bleackley) contain as only significant homology cAMP response elements. These findings are consistent with a tight control and regulation of P1, which appears to be distinct from that of granzymes.",
author = "Lichtenheld, {Mathias G} and Podack, {E. R.}",
year = "1989",
month = "12",
day = "1",
language = "English",
volume = "143",
pages = "4267--4274",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "12",

}

TY - JOUR

T1 - Structure of the human perforin gene. A simple gene organization with interesting potential regulatory sequences

AU - Lichtenheld, Mathias G

AU - Podack, E. R.

PY - 1989/12/1

Y1 - 1989/12/1

N2 - We have cloned the human perforin (P1) gene and sequenced 6.2-kb genomic DNA, containing 1.4-kb 5'-flanking region, the 5' untranslated region, the complete coding region and the beginning of the 3' untranslated region. The P1 gene including at least 95-bp 3' untranslated region is organized in only three exons: the first exon (97 bp) contains all but four nucleotides of the 5' untranslated region and was determined by primer extension and S1 nuclease mapping. This exon is separated by 1.7 kb from the second exon containing the remaining (4 bp) 5' untranslated region, the leader peptide and the N-terminal region of P1 up to - but not including - the C9 homologous region. The third exon is separated by a 1.2-kb intron and contains the remainder of the molecule, including at least 90 bp of the 3' untranslated region. This simple gene organization differs from that of the more complicated C9 gene. Because of the unusual intron in the 5' untranslated sequence the transcription initiation (cap) site is located almost 1.8 kb upstream of the ATG start signal. The more immediate 5' flanking sequence contains a CCAAT and GC box but lacks other known promoter elements. Instead, we find three different sequence repeats. One of them, a hexanucleotide sequence with the consensus GCCCTG of unknown significance occurs 19 times within a stretch of 240 bp. Further upstream we localized sequences homologous to the following enhancer and promoter elements: c-fos proto-oncogen, IFN-γ and phorbol ester response elements, five cAMP response elements, and three motifs corresponding to general inducer elements. In addition, a sequence conserved in the 5' flanking region of several T cell genes was identified. The 5' flanking regions of P1, CCP1 (granzyme B) and CCP2 (granzyme C) (kindly provided by Dr. Bleackley) contain as only significant homology cAMP response elements. These findings are consistent with a tight control and regulation of P1, which appears to be distinct from that of granzymes.

AB - We have cloned the human perforin (P1) gene and sequenced 6.2-kb genomic DNA, containing 1.4-kb 5'-flanking region, the 5' untranslated region, the complete coding region and the beginning of the 3' untranslated region. The P1 gene including at least 95-bp 3' untranslated region is organized in only three exons: the first exon (97 bp) contains all but four nucleotides of the 5' untranslated region and was determined by primer extension and S1 nuclease mapping. This exon is separated by 1.7 kb from the second exon containing the remaining (4 bp) 5' untranslated region, the leader peptide and the N-terminal region of P1 up to - but not including - the C9 homologous region. The third exon is separated by a 1.2-kb intron and contains the remainder of the molecule, including at least 90 bp of the 3' untranslated region. This simple gene organization differs from that of the more complicated C9 gene. Because of the unusual intron in the 5' untranslated sequence the transcription initiation (cap) site is located almost 1.8 kb upstream of the ATG start signal. The more immediate 5' flanking sequence contains a CCAAT and GC box but lacks other known promoter elements. Instead, we find three different sequence repeats. One of them, a hexanucleotide sequence with the consensus GCCCTG of unknown significance occurs 19 times within a stretch of 240 bp. Further upstream we localized sequences homologous to the following enhancer and promoter elements: c-fos proto-oncogen, IFN-γ and phorbol ester response elements, five cAMP response elements, and three motifs corresponding to general inducer elements. In addition, a sequence conserved in the 5' flanking region of several T cell genes was identified. The 5' flanking regions of P1, CCP1 (granzyme B) and CCP2 (granzyme C) (kindly provided by Dr. Bleackley) contain as only significant homology cAMP response elements. These findings are consistent with a tight control and regulation of P1, which appears to be distinct from that of granzymes.

UR - http://www.scopus.com/inward/record.url?scp=0024822421&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024822421&partnerID=8YFLogxK

M3 - Article

C2 - 2480391

AN - SCOPUS:0024822421

VL - 143

SP - 4267

EP - 4274

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 12

ER -