Identification of the promoter, genomic structure, and mouse ortholog of LGI1

Robert P.T. Somerville, Olga Chernova, Siqing Liu, Yigal Shoshan, John Kenneth Cowell

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The human LGI1 gene is a leucine-rich, repeat-containing gene that was cloned from the t(10:19) breakpoint of the T98G glioblastoma cell line. The LGI1 gene maps to 10q24, a region of peak LOH in malignant gliomas, and is inactivated during the transition from low to high-grade brain tumors. Here we report detailed studies of the genomic structure of the LGI1 gene and its promoter. We have also cloned and characterized the mouse lgil gene, which is 97% homologous to the human gene at the amino acid level and 91% homologous at the nucleotide level. LGI1 contains 8 exons, where each of the four leucine-rich repeat units is contained in an individual 72-bp exon. The cysteine-rich regions flanking the LRR and the single trans-membrane domain do not occupy individual exons. Approximately 5-kb of the genomic region 5' to LGI1 was sequenced, but conventional CAAT and TATA motifs were not present within this sequence. A 597-bp fragment of this 5' sequence was cloned upstream of a promoterless luciferase gene and was shown to be sufficient to drive transcription. SSCP analysis of the coding region of LGI1 in 20 glioblastomas and five cell lines did not reveal any mutations. Because LGI1 expression is considerably downregulated in gliomas, we also investigated whether this was owing to changes in the methylation status of the promoter. Southern blot analysis and 5-azacytidine treatment did not show any appreciable difference in methylation status between normal brain and glioblastomas.

Original languageEnglish (US)
Pages (from-to)622-627
Number of pages6
JournalMammalian Genome
Volume11
Issue number8
DOIs
StatePublished - Jan 1 2000
Externally publishedYes

Fingerprint

Genes
Glioblastoma
Exons
Leucine
Glioma
Methylation
Azacitidine
Single-Stranded Conformational Polymorphism
Cell Line
Southern Blotting
Luciferases
Brain Neoplasms
Cysteine
Down-Regulation
Nucleotides
Amino Acids
Mutation
Membranes
Brain

ASJC Scopus subject areas

  • Genetics

Cite this

Identification of the promoter, genomic structure, and mouse ortholog of LGI1. / Somerville, Robert P.T.; Chernova, Olga; Liu, Siqing; Shoshan, Yigal; Cowell, John Kenneth.

In: Mammalian Genome, Vol. 11, No. 8, 01.01.2000, p. 622-627.

Research output: Contribution to journalArticle

Somerville, Robert P.T. ; Chernova, Olga ; Liu, Siqing ; Shoshan, Yigal ; Cowell, John Kenneth. / Identification of the promoter, genomic structure, and mouse ortholog of LGI1. In: Mammalian Genome. 2000 ; Vol. 11, No. 8. pp. 622-627.
@article{fc243f8835554018a5c52d02dfe8cd32,
title = "Identification of the promoter, genomic structure, and mouse ortholog of LGI1",
abstract = "The human LGI1 gene is a leucine-rich, repeat-containing gene that was cloned from the t(10:19) breakpoint of the T98G glioblastoma cell line. The LGI1 gene maps to 10q24, a region of peak LOH in malignant gliomas, and is inactivated during the transition from low to high-grade brain tumors. Here we report detailed studies of the genomic structure of the LGI1 gene and its promoter. We have also cloned and characterized the mouse lgil gene, which is 97{\%} homologous to the human gene at the amino acid level and 91{\%} homologous at the nucleotide level. LGI1 contains 8 exons, where each of the four leucine-rich repeat units is contained in an individual 72-bp exon. The cysteine-rich regions flanking the LRR and the single trans-membrane domain do not occupy individual exons. Approximately 5-kb of the genomic region 5' to LGI1 was sequenced, but conventional CAAT and TATA motifs were not present within this sequence. A 597-bp fragment of this 5' sequence was cloned upstream of a promoterless luciferase gene and was shown to be sufficient to drive transcription. SSCP analysis of the coding region of LGI1 in 20 glioblastomas and five cell lines did not reveal any mutations. Because LGI1 expression is considerably downregulated in gliomas, we also investigated whether this was owing to changes in the methylation status of the promoter. Southern blot analysis and 5-azacytidine treatment did not show any appreciable difference in methylation status between normal brain and glioblastomas.",
author = "Somerville, {Robert P.T.} and Olga Chernova and Siqing Liu and Yigal Shoshan and Cowell, {John Kenneth}",
year = "2000",
month = "1",
day = "1",
doi = "10.1007/s0033500101280",
language = "English (US)",
volume = "11",
pages = "622--627",
journal = "Mammalian Genome",
issn = "0938-8990",
publisher = "Springer New York",
number = "8",

}

TY - JOUR

T1 - Identification of the promoter, genomic structure, and mouse ortholog of LGI1

AU - Somerville, Robert P.T.

AU - Chernova, Olga

AU - Liu, Siqing

AU - Shoshan, Yigal

AU - Cowell, John Kenneth

PY - 2000/1/1

Y1 - 2000/1/1

N2 - The human LGI1 gene is a leucine-rich, repeat-containing gene that was cloned from the t(10:19) breakpoint of the T98G glioblastoma cell line. The LGI1 gene maps to 10q24, a region of peak LOH in malignant gliomas, and is inactivated during the transition from low to high-grade brain tumors. Here we report detailed studies of the genomic structure of the LGI1 gene and its promoter. We have also cloned and characterized the mouse lgil gene, which is 97% homologous to the human gene at the amino acid level and 91% homologous at the nucleotide level. LGI1 contains 8 exons, where each of the four leucine-rich repeat units is contained in an individual 72-bp exon. The cysteine-rich regions flanking the LRR and the single trans-membrane domain do not occupy individual exons. Approximately 5-kb of the genomic region 5' to LGI1 was sequenced, but conventional CAAT and TATA motifs were not present within this sequence. A 597-bp fragment of this 5' sequence was cloned upstream of a promoterless luciferase gene and was shown to be sufficient to drive transcription. SSCP analysis of the coding region of LGI1 in 20 glioblastomas and five cell lines did not reveal any mutations. Because LGI1 expression is considerably downregulated in gliomas, we also investigated whether this was owing to changes in the methylation status of the promoter. Southern blot analysis and 5-azacytidine treatment did not show any appreciable difference in methylation status between normal brain and glioblastomas.

AB - The human LGI1 gene is a leucine-rich, repeat-containing gene that was cloned from the t(10:19) breakpoint of the T98G glioblastoma cell line. The LGI1 gene maps to 10q24, a region of peak LOH in malignant gliomas, and is inactivated during the transition from low to high-grade brain tumors. Here we report detailed studies of the genomic structure of the LGI1 gene and its promoter. We have also cloned and characterized the mouse lgil gene, which is 97% homologous to the human gene at the amino acid level and 91% homologous at the nucleotide level. LGI1 contains 8 exons, where each of the four leucine-rich repeat units is contained in an individual 72-bp exon. The cysteine-rich regions flanking the LRR and the single trans-membrane domain do not occupy individual exons. Approximately 5-kb of the genomic region 5' to LGI1 was sequenced, but conventional CAAT and TATA motifs were not present within this sequence. A 597-bp fragment of this 5' sequence was cloned upstream of a promoterless luciferase gene and was shown to be sufficient to drive transcription. SSCP analysis of the coding region of LGI1 in 20 glioblastomas and five cell lines did not reveal any mutations. Because LGI1 expression is considerably downregulated in gliomas, we also investigated whether this was owing to changes in the methylation status of the promoter. Southern blot analysis and 5-azacytidine treatment did not show any appreciable difference in methylation status between normal brain and glioblastomas.

UR - http://www.scopus.com/inward/record.url?scp=0038285465&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038285465&partnerID=8YFLogxK

U2 - 10.1007/s0033500101280

DO - 10.1007/s0033500101280

M3 - Article

C2 - 10920229

AN - SCOPUS:0038285465

VL - 11

SP - 622

EP - 627

JO - Mammalian Genome

JF - Mammalian Genome

SN - 0938-8990

IS - 8

ER -