Identification of two novel mutations and of a novel critical region in the KRIT1 gene

Vito Guarnieri, Lucia A. Muscarella, Rosina Amoroso, Alessandro Quattrone, Massimo E. Abate, Michelina Coco, Domenico Catapano, Vincenzo A. D'Angelo, Leopoldo Zelante, Leonardo D'Agruma

Research output: Contribution to journalArticle

Abstract

Cerebral cavernous malformations (CCMs) represent a common autosomal dominant disorder that predisposes patients to hemorrhagic strokes and focal neurological signs. Mutations in three genes (KRIT1, MGC4607, and PDCD10) have been associated with CCMs. We investigated the role of two new mutations in the KRIT1 gene in two Italian families affected by CCMs. Whole blood DNA was extracted and the mutations were detected after polymerase chain reaction (PCR), denaturing high-performance liquid chromatography screening, and sequencing of the coding regions of the three CCMs-associated genes. Total RNA was extracted, and the KRIT1 cDNA was sequenced and subsequently subjected to real-time quantitative PCR in order to examine the translational outcome of each genomic mutation. A novel splicing acceptor site deletion of the exon 14 in one family and an intronic nucleotide change close to the exon 19 in the other one were identified, both in the KRIT1 gene. These mutations were proven to alter the correct splicing mechanism, resulting, respectively, in a truncated protein of 432 amino acids and in a protein lacking an internal segment. We report two novel cases of splicing affecting genomic variants, suggesting a careful reanalysis of previously identified splice site variations in KRIT1 to look for their possible causative roles of similar missplicing events and their consequent involvement in the pathogenesis of CCMs. Moreover, our genotype-phenotype functional correlation suggests that the C-terminal portion of the KRIT1 protein is likely to contain a short, previously unrecognized segment necessary for its activity.

Original languageEnglish
Pages (from-to)29-37
Number of pages9
JournalNeurogenetics
Volume8
Issue number1
DOIs
Publication statusPublished - Jan 2007

Fingerprint

Central Nervous System Cavernous Hemangioma
Mutation
Genes
Exons
Proteins
Genetic Association Studies
Real-Time Polymerase Chain Reaction
Nucleotides
Complementary DNA
Stroke
High Pressure Liquid Chromatography
RNA
Amino Acids
Polymerase Chain Reaction
DNA

Keywords

  • Genetics
  • Mutation
  • RTqPCR
  • SpaGV
  • Vascular malformations

ASJC Scopus subject areas

  • Genetics(clinical)
  • Neuroscience(all)

Cite this

Identification of two novel mutations and of a novel critical region in the KRIT1 gene. / Guarnieri, Vito; Muscarella, Lucia A.; Amoroso, Rosina; Quattrone, Alessandro; Abate, Massimo E.; Coco, Michelina; Catapano, Domenico; D'Angelo, Vincenzo A.; Zelante, Leopoldo; D'Agruma, Leonardo.

In: Neurogenetics, Vol. 8, No. 1, 01.2007, p. 29-37.

Research output: Contribution to journalArticle

Guarnieri, Vito ; Muscarella, Lucia A. ; Amoroso, Rosina ; Quattrone, Alessandro ; Abate, Massimo E. ; Coco, Michelina ; Catapano, Domenico ; D'Angelo, Vincenzo A. ; Zelante, Leopoldo ; D'Agruma, Leonardo. / Identification of two novel mutations and of a novel critical region in the KRIT1 gene. In: Neurogenetics. 2007 ; Vol. 8, No. 1. pp. 29-37.
@article{6bafa6ad0e1448b99b7dd43256a9f52c,
title = "Identification of two novel mutations and of a novel critical region in the KRIT1 gene",
abstract = "Cerebral cavernous malformations (CCMs) represent a common autosomal dominant disorder that predisposes patients to hemorrhagic strokes and focal neurological signs. Mutations in three genes (KRIT1, MGC4607, and PDCD10) have been associated with CCMs. We investigated the role of two new mutations in the KRIT1 gene in two Italian families affected by CCMs. Whole blood DNA was extracted and the mutations were detected after polymerase chain reaction (PCR), denaturing high-performance liquid chromatography screening, and sequencing of the coding regions of the three CCMs-associated genes. Total RNA was extracted, and the KRIT1 cDNA was sequenced and subsequently subjected to real-time quantitative PCR in order to examine the translational outcome of each genomic mutation. A novel splicing acceptor site deletion of the exon 14 in one family and an intronic nucleotide change close to the exon 19 in the other one were identified, both in the KRIT1 gene. These mutations were proven to alter the correct splicing mechanism, resulting, respectively, in a truncated protein of 432 amino acids and in a protein lacking an internal segment. We report two novel cases of splicing affecting genomic variants, suggesting a careful reanalysis of previously identified splice site variations in KRIT1 to look for their possible causative roles of similar missplicing events and their consequent involvement in the pathogenesis of CCMs. Moreover, our genotype-phenotype functional correlation suggests that the C-terminal portion of the KRIT1 protein is likely to contain a short, previously unrecognized segment necessary for its activity.",
keywords = "Genetics, Mutation, RTqPCR, SpaGV, Vascular malformations",
author = "Vito Guarnieri and Muscarella, {Lucia A.} and Rosina Amoroso and Alessandro Quattrone and Abate, {Massimo E.} and Michelina Coco and Domenico Catapano and D'Angelo, {Vincenzo A.} and Leopoldo Zelante and Leonardo D'Agruma",
year = "2007",
month = "1",
doi = "10.1007/s10048-006-0056-y",
language = "English",
volume = "8",
pages = "29--37",
journal = "Neurogenetics",
issn = "1364-6745",
publisher = "Springer Verlag",
number = "1",

}

TY - JOUR

T1 - Identification of two novel mutations and of a novel critical region in the KRIT1 gene

AU - Guarnieri, Vito

AU - Muscarella, Lucia A.

AU - Amoroso, Rosina

AU - Quattrone, Alessandro

AU - Abate, Massimo E.

AU - Coco, Michelina

AU - Catapano, Domenico

AU - D'Angelo, Vincenzo A.

AU - Zelante, Leopoldo

AU - D'Agruma, Leonardo

PY - 2007/1

Y1 - 2007/1

N2 - Cerebral cavernous malformations (CCMs) represent a common autosomal dominant disorder that predisposes patients to hemorrhagic strokes and focal neurological signs. Mutations in three genes (KRIT1, MGC4607, and PDCD10) have been associated with CCMs. We investigated the role of two new mutations in the KRIT1 gene in two Italian families affected by CCMs. Whole blood DNA was extracted and the mutations were detected after polymerase chain reaction (PCR), denaturing high-performance liquid chromatography screening, and sequencing of the coding regions of the three CCMs-associated genes. Total RNA was extracted, and the KRIT1 cDNA was sequenced and subsequently subjected to real-time quantitative PCR in order to examine the translational outcome of each genomic mutation. A novel splicing acceptor site deletion of the exon 14 in one family and an intronic nucleotide change close to the exon 19 in the other one were identified, both in the KRIT1 gene. These mutations were proven to alter the correct splicing mechanism, resulting, respectively, in a truncated protein of 432 amino acids and in a protein lacking an internal segment. We report two novel cases of splicing affecting genomic variants, suggesting a careful reanalysis of previously identified splice site variations in KRIT1 to look for their possible causative roles of similar missplicing events and their consequent involvement in the pathogenesis of CCMs. Moreover, our genotype-phenotype functional correlation suggests that the C-terminal portion of the KRIT1 protein is likely to contain a short, previously unrecognized segment necessary for its activity.

AB - Cerebral cavernous malformations (CCMs) represent a common autosomal dominant disorder that predisposes patients to hemorrhagic strokes and focal neurological signs. Mutations in three genes (KRIT1, MGC4607, and PDCD10) have been associated with CCMs. We investigated the role of two new mutations in the KRIT1 gene in two Italian families affected by CCMs. Whole blood DNA was extracted and the mutations were detected after polymerase chain reaction (PCR), denaturing high-performance liquid chromatography screening, and sequencing of the coding regions of the three CCMs-associated genes. Total RNA was extracted, and the KRIT1 cDNA was sequenced and subsequently subjected to real-time quantitative PCR in order to examine the translational outcome of each genomic mutation. A novel splicing acceptor site deletion of the exon 14 in one family and an intronic nucleotide change close to the exon 19 in the other one were identified, both in the KRIT1 gene. These mutations were proven to alter the correct splicing mechanism, resulting, respectively, in a truncated protein of 432 amino acids and in a protein lacking an internal segment. We report two novel cases of splicing affecting genomic variants, suggesting a careful reanalysis of previously identified splice site variations in KRIT1 to look for their possible causative roles of similar missplicing events and their consequent involvement in the pathogenesis of CCMs. Moreover, our genotype-phenotype functional correlation suggests that the C-terminal portion of the KRIT1 protein is likely to contain a short, previously unrecognized segment necessary for its activity.

KW - Genetics

KW - Mutation

KW - RTqPCR

KW - SpaGV

KW - Vascular malformations

UR - http://www.scopus.com/inward/record.url?scp=33845668391&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33845668391&partnerID=8YFLogxK

U2 - 10.1007/s10048-006-0056-y

DO - 10.1007/s10048-006-0056-y

M3 - Article

C2 - 17043900

AN - SCOPUS:33845668391

VL - 8

SP - 29

EP - 37

JO - Neurogenetics

JF - Neurogenetics

SN - 1364-6745

IS - 1

ER -