Differential splicing of Pneumocystis carinii f. sp. carinii inosine 5′-monophosphate dehydrogenase pre-mRNA

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Inosine 5′-monophosphate dehydrogenase (IMPDH) is a rate-limiting enzyme in guanine nucleotide metabolism that has drawn attention as a drug target in several organisms. Pneumocystis carinii f. sp. carinii IMPDH mRNA (GeneBank Accession No: U42442) previously identified from cultured organisms yielded a predicted amino acid sequence about 70 amino acids shorter at the amino terminus than IMPDH from other species. Recent research has shown that the amino terminal region is important for enzyme activity, suggesting that the previous putative P. carinii IMPDH might not represent full length, functional enzyme. To test this hypothesis, RT-PCR was performed with total RNA isolated from P. carinii f. sp. carinii. Three IMPDH splicing variants were found and splicing preference was observed: P. carinii isolated from infected rat lung contained primarily splicing variant one (introns two and four deleted), but organisms from spinner flask culture contained primarily splicing variant three (all four introns deleted). Importantly, splicing variant one (GeneBank Accession No: AF196975) contained an open reading frame for 529 amino acids, a size comparable to that of other eukaryotic IMPDH forms. The other variants contained the same open reading frame (454 amino acids) previously reported. Sequence analysis and complementation studies suggest variant one represents the full length, catalytically active form of P. carinii IMPDH. The differential splicing of the enzyme may reflect a mechanism by which the organism regulates the expression of IMPDH in response to environmental stresses.

Original languageEnglish
Pages (from-to)151-158
Number of pages8
JournalGene
Volume263
Issue number1-2
DOIs
StatePublished - Jan 24 2001

Fingerprint

IMP Dehydrogenase
Pneumocystis carinii
RNA Precursors
Enzymes
Amino Acids
Introns
Open Reading Frames
Guanine Nucleotides
Sequence Analysis
Amino Acid Sequence
RNA
Polymerase Chain Reaction
Lung
Messenger RNA

Keywords

  • Alternative splicing
  • Drug targets
  • Introns
  • Opportunistic infections
  • Purine metabolism

ASJC Scopus subject areas

  • Genetics

Cite this

Differential splicing of Pneumocystis carinii f. sp. carinii inosine 5′-monophosphate dehydrogenase pre-mRNA. / Ye, Dongjiu; Lee, Chao-Hung; Queener, Sherry.

In: Gene, Vol. 263, No. 1-2, 24.01.2001, p. 151-158.

Research output: Contribution to journalArticle

@article{3cf95a1503ba4f819f02248aa856a9d6,
title = "Differential splicing of Pneumocystis carinii f. sp. carinii inosine 5′-monophosphate dehydrogenase pre-mRNA",
abstract = "Inosine 5′-monophosphate dehydrogenase (IMPDH) is a rate-limiting enzyme in guanine nucleotide metabolism that has drawn attention as a drug target in several organisms. Pneumocystis carinii f. sp. carinii IMPDH mRNA (GeneBank Accession No: U42442) previously identified from cultured organisms yielded a predicted amino acid sequence about 70 amino acids shorter at the amino terminus than IMPDH from other species. Recent research has shown that the amino terminal region is important for enzyme activity, suggesting that the previous putative P. carinii IMPDH might not represent full length, functional enzyme. To test this hypothesis, RT-PCR was performed with total RNA isolated from P. carinii f. sp. carinii. Three IMPDH splicing variants were found and splicing preference was observed: P. carinii isolated from infected rat lung contained primarily splicing variant one (introns two and four deleted), but organisms from spinner flask culture contained primarily splicing variant three (all four introns deleted). Importantly, splicing variant one (GeneBank Accession No: AF196975) contained an open reading frame for 529 amino acids, a size comparable to that of other eukaryotic IMPDH forms. The other variants contained the same open reading frame (454 amino acids) previously reported. Sequence analysis and complementation studies suggest variant one represents the full length, catalytically active form of P. carinii IMPDH. The differential splicing of the enzyme may reflect a mechanism by which the organism regulates the expression of IMPDH in response to environmental stresses.",
keywords = "Alternative splicing, Drug targets, Introns, Opportunistic infections, Purine metabolism",
author = "Dongjiu Ye and Chao-Hung Lee and Sherry Queener",
year = "2001",
month = "1",
day = "24",
doi = "10.1016/S0378-1119(00)00577-1",
language = "English",
volume = "263",
pages = "151--158",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Differential splicing of Pneumocystis carinii f. sp. carinii inosine 5′-monophosphate dehydrogenase pre-mRNA

AU - Ye, Dongjiu

AU - Lee, Chao-Hung

AU - Queener, Sherry

PY - 2001/1/24

Y1 - 2001/1/24

N2 - Inosine 5′-monophosphate dehydrogenase (IMPDH) is a rate-limiting enzyme in guanine nucleotide metabolism that has drawn attention as a drug target in several organisms. Pneumocystis carinii f. sp. carinii IMPDH mRNA (GeneBank Accession No: U42442) previously identified from cultured organisms yielded a predicted amino acid sequence about 70 amino acids shorter at the amino terminus than IMPDH from other species. Recent research has shown that the amino terminal region is important for enzyme activity, suggesting that the previous putative P. carinii IMPDH might not represent full length, functional enzyme. To test this hypothesis, RT-PCR was performed with total RNA isolated from P. carinii f. sp. carinii. Three IMPDH splicing variants were found and splicing preference was observed: P. carinii isolated from infected rat lung contained primarily splicing variant one (introns two and four deleted), but organisms from spinner flask culture contained primarily splicing variant three (all four introns deleted). Importantly, splicing variant one (GeneBank Accession No: AF196975) contained an open reading frame for 529 amino acids, a size comparable to that of other eukaryotic IMPDH forms. The other variants contained the same open reading frame (454 amino acids) previously reported. Sequence analysis and complementation studies suggest variant one represents the full length, catalytically active form of P. carinii IMPDH. The differential splicing of the enzyme may reflect a mechanism by which the organism regulates the expression of IMPDH in response to environmental stresses.

AB - Inosine 5′-monophosphate dehydrogenase (IMPDH) is a rate-limiting enzyme in guanine nucleotide metabolism that has drawn attention as a drug target in several organisms. Pneumocystis carinii f. sp. carinii IMPDH mRNA (GeneBank Accession No: U42442) previously identified from cultured organisms yielded a predicted amino acid sequence about 70 amino acids shorter at the amino terminus than IMPDH from other species. Recent research has shown that the amino terminal region is important for enzyme activity, suggesting that the previous putative P. carinii IMPDH might not represent full length, functional enzyme. To test this hypothesis, RT-PCR was performed with total RNA isolated from P. carinii f. sp. carinii. Three IMPDH splicing variants were found and splicing preference was observed: P. carinii isolated from infected rat lung contained primarily splicing variant one (introns two and four deleted), but organisms from spinner flask culture contained primarily splicing variant three (all four introns deleted). Importantly, splicing variant one (GeneBank Accession No: AF196975) contained an open reading frame for 529 amino acids, a size comparable to that of other eukaryotic IMPDH forms. The other variants contained the same open reading frame (454 amino acids) previously reported. Sequence analysis and complementation studies suggest variant one represents the full length, catalytically active form of P. carinii IMPDH. The differential splicing of the enzyme may reflect a mechanism by which the organism regulates the expression of IMPDH in response to environmental stresses.

KW - Alternative splicing

KW - Drug targets

KW - Introns

KW - Opportunistic infections

KW - Purine metabolism

UR - http://www.scopus.com/inward/record.url?scp=0035941463&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035941463&partnerID=8YFLogxK

U2 - 10.1016/S0378-1119(00)00577-1

DO - 10.1016/S0378-1119(00)00577-1

M3 - Article

VL - 263

SP - 151

EP - 158

JO - Gene

JF - Gene

SN - 0378-1119

IS - 1-2

ER -