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J H Kinnaird

Publications and source records attributed to J H Kinnaird.

11 recordsLinked to original sources
PubMed2004

EtCRK2, a cyclin-dependent kinase gene expressed during the sexual and asexual phases of the Eimeria tenella life cycle.

EtCRK2, a cyclin-dependent kinase from the coccidian parasite, Eimeria tenella is closely related to eukaryotic cyclin-dependent kinases that regulate progression of the cell cycle and to several cyclin-dependent kinases identified in the Apicomplexa. Northern blot analyses revealed that EtCRK2 is transcribed during both asexual (first-generation schizogony) and sexual (oocyst sporulation) replicative phases of the parasite life cycle. In addition, it appears to be transcriptionally regulated during meiosis. Recombinant EtCRK2 produced in Escherichia coli has kinase activity which is significantly stimulated by the addition of vertebrate cyclin A. This cyclin-dependent kinase may play a significant role in regulating critical cell cycle events during both asexual proliferation and sexual development of the parasite.

Amino Acid Sequence↗
PubMed1999

Ribonucleotide reductase: A new target for antiparasite therapies.

New treatments are required urgently for the control of parasitic protozoan diseases of humans and animals. One approach is the development of subunit vaccines; the other focuses on identifying and exploiting specific differences in essential functions between the host and parasite. One enzyme currently attracting attention is ribonucleotide reductase, an essential component in the biosynthesis of DNA. In this article, Geoffrey Ingram and Jane Kinnaird examine differences between the host and parasite enzymes and assess possible means of therapeutic intervention.

Amino Acid Sequence↗
PubMed1996

The isolation and characterization of genomic and cDNA clones coding for a cdc2-related kinase (ThCRK2) from the bovine protozoan parasite Theileria.

The tick-transmitted protozoan parasites Theileria annulata and Theileria parva are important intracellular pathogens of domestic cattle in tropical and subtropical regions. Proliferative phases take place within both lymphocytes and erythrocytes. The lymphocyte is stimulated to enter the cell cycle by the parasite and the multinucleate parasite can establish a state in which karyokinesis and cytokinesis occur in phase with the host cell. The link between parasite nuclear division and cytokinesis is altered during the formation of merozoites (a non-dividing, invasive, extracellular stage). These features imply a high degree of control over parasite nuclear division and cytokinesis. Two different approaches have been used to identify clones from both species which are extremely highly conserved homologues. These encode a cdc2-related kinase which is > 60% identical to eukaryotic cyclin-dependent kinases of the p34cdc2/p32CDK2 subfamily. There is typical conservation of kinase domains, implying an in vivo protein kinase activity for the polypeptide. The PSTAIRE region, implicated in cyclin binding, is well conserved suggesting that ThCRK2 will bind cyclin molecules closely related to the eukaryotic A/B-type cyclins. However, there is divergence in certain key motifs potentially associated with binding of molecules that regulate the activity of the kinase. Expression patterns of RNA and protein indicate that ThCRK2 is likely to function in all dividing stages of the parasite and, taken together, the results point to a central role in the regulation of nuclear division.

Amino Acid Sequence↗
PubMed1993

A novel CDC2-related protein kinase from Leishmania mexicana, LmmCRK1, is post-translationally regulated during the life cycle.

The p34CDC2 protein kinase is a key component in the regulation of the eukaryotic cell cycle. We have isolated from the protozoan parasite Leishmania mexicana mexicana a CDC2-related kinase gene (Lmmcrk1) encoding a 34-kDa protein kinase (lmmCRK1) which has 56% amino acid identity with the human CDC2 and contains a PCTAIR motif in place of the highly conserved PSTAIR box. lmmCRK1 was detected in all life cycle stages at comparable levels, yet its histone H1 kinase activity was detected in only the promastigote form, indicating that its activity is stage-regulated at a post-translational level. lmmCRK1 did not bind p13suc1 beads and Lmmcrk1 was unable to complement a fission yeast temperature-sensitive cdc2 mutant. These data suggest that Lmmcrk1 is unlikely to be the functional L. mexicana cdc2 homologue. A distinct histone H1 kinase activity that binds p13suc1 beads (SBCRK) was also detected, with activity that correlated with the division status of the developmental forms of the parasite, being present in the dividing stages of the parasite and absent in nondividing metacyclic forms. SBCRK is a candidate for the functional CDC2 homologue, but it does not react with an anti-PSTAIR monoclonal antibody on Western blots when eluted from p13suc1 beads, indicating a divergent PSTAIR box. These data suggest that a family of CDC2-related protein kinases are present in Leishmania. Some share sequence and biochemical properties with CDC2, but significant differences also exist, possibly reflecting the evolutionary distance between Leishmania and higher eukaryotes.

Amino Acid Sequence↗
PubMed1992

Alternative modes of mRNA processing in a 3' splice site mutant of Neurospora crassa.

The am8 mutant of Neurospora crassa is shown to have a double base-pair change, GTG for the normal TAG, at the 3' end of the second intron of the am (NADP-specific glutamate dehydrogenase, GDH) gene. The greater part of the mutant am transcript accumulates as two fragments hybridising to probes for sequences respectively upstream and downstream of the 5' end of the intron. Two processed transcripts approximating to normal full length mRNA were identified. In one the second intron was intact; in the other the second intron was spliced out through the use of an AAG sequence, 20 base-pairs into the third exon, as a 3' acceptor site. A GAG sequence, only four base-pairs downstream from the normal acceptor site, does not appear to be used.

Base Sequence↗
PubMed1991

An apparent rare-codon effect on the rate of translation of a Neurospora gene.

As the result of two mutually compensating frameshift mutations, three successive codons with third-position A were generated in the Neurospora crassa am (NADP-specific glutamate dehydrogenase: GDH) gene. These codons do not occur at all elsewhere in the gene and only infrequently in other highly expressed Neurospora genes. The double-frameshift strain produces only 25 to 35% of the normal level of GDH, whether measured as enzyme activity or as immunoprecipitable protein, but its level of GDH mRNA is normal. Although the modified enzyme is somewhat more heat-sensitive than the wild-type in vitro, its stability in vivo was found to be indistinguishable from that of the wild-type. It is concluded that the introduction of consecutive rare codons reduces the efficiency of translation of the mRNA. The possible mechanisms of such an effect are discussed.

Amino Acid Sequence↗
PubMed1986

Sequencing studies of ICR-170 mutagenic specificity in the am (NADP-specific glutamate dehydrogenase) gene of Neurospora crassa.

The acridine half-mustard ICR-170-induced reversion of the mutant am15, which has a single base-pair deletion, at a frequency of between 9 and 28 X 10(-6). In each of three classes of revertants, the mutagen had induced the insertion of a -G- -C- base pair at a -G-G- -C-C- site. The mutant am6, which has a single base pair insertion, is known to be revertible, with UV light, by deletion of a -G- -C- base pair at a -G-G-G- -C-C-C- site. This mutant reverted with ICR-170 at a frequency of 0.1 X 10(-6). These results show that ICR-170 is able to induce addition frameshifts in Neurospora crassa within short, monotonous runs of G:C base pairs, but indicate a lack of deletion activity at such sequences.

Amino Acid Sequence↗
PubMed1983

The complete nucleotide sequence of the Neurospora crassa am (NADP-specific glutamate dehydrogenase) gene.

The complete nucleotide sequence of a 2.7-kb genomic fragment, containing the Neurospora crassa am [NADP-specific glutamate dehydrogenase (GDH)] gene, has been determined. The transcription initiation and polyadenylation sites have been defined by S1 mapping. There are at least four initiation sites between 35 and 60 bases downstream of a TATAAA sequence. The single polyadenylation site is immediately downstream of a six-nucleotide sequence which is present in the corresponding position in the N. crassa gene (pl) coding for the imported proteolipid component of mitochondrial ATP synthase [Viebrock et al. Embo J. 1 (1982) 565-571] the initiation sites also show some sequence similarity between am and pl. There are two introns, of 66 and 61 bases respectively; they interrupt sequences that are conserved (highly so in the case of the second intron) between Neurospora and Escherichia coli. As well as showing typical consensus splicing sequences, the two introns show some striking internal similarities to each other. The am gene resembles the pl gene in its strongly selective use of codons (pyrimidines preferred to purines and guanine to adenine in third positions). The coding sequence here reported requires four corrections to be made in the previously published amino acid sequence of Neurospora GDH.

Amino Acid Sequence↗
PubMed1982

Cloning of the am (glutamate dehydrogenase) gene of Neurospora crassa through the use of a synthetic DNA probe.

In a previous study the alteration in the amino acid sequence of Neurospora crassa NADP-specific glutamate dehydrogenase (GDH) resulting from two mutually compensating frameshift mutations was used to deduce the first 17 nucleotides of the coding sequence of the am gene. In the work reported here, a synthetic 17-mer corresponding to the deduced sequence was shown to hybridize strongly to a 9-kb HindIII fragment from N. crassa wild-type DNA but not to any corresponding fragment from the DNA of a mutant strain known to be deleted for most or all of the gene. Wild-type HindIII fragments were fractionated for size and a fraction centering around 9 kb was cloned in vector lambda L47. Two clones carrying the strongly hybridizing fragment were identified. The hybridization to the 17-mer was localized within a 2.7-kb BamHI fragment and, within this, to a 700-bp BamHI-Bg/II subfragment. 5' end-labelled polyadenylated RNA isolated from wild-type mycelium hybridized to the 2.7-kb BamHI fragment and not appreciably to flanking fragments. The partial sequence analysis of the BamHI-Bg/II fragment has confirmed that the 17-mer probe matches the coding sequence at the 5' end of the gene and has also revealed an intervening sequence 67 bp in length, interrupting codon 15. Both the 9-kb HindIII fragment and the 2.7-kb BamHI fragment have been shown to be capable of transforming the deletion mutant to prototrophy and ability to produce GDH. Analysis of one transformant showed that the am gene was integrated, together with a part of the long arm of the lambda vector, at an unusual locus. This transformant, in which the am gene does not show its normal linkage to the linkage group 5 marker inl, was found to produce GDH to about 20% of the normal level.

Bacteriophage lambda↗