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H Masai

Publications and source records attributed to H Masai.

At least 55 records · Page 3Linked to original sources
PubMed1991

Identification of eleven single-strand initiation sequences (ssi) for priming of DNA replication in the F, R6K, R100 and ColE2 plasmids.

Based on the ability to complement the poor growth of an M13 phage derivative lacking the complementary strand origin, eleven single-strand initiation sequences (ssi) for DNA replication are identified in the F, R6K, R100 and ColE2 plasmids. Six of them were from F, two from near the gamma and alpha origins (ori) of R6K, two from the vicinity of the basic replicon of R100 and one from near the ori of ColE2. They can be classified into two groups based on the morphology of the plaques and the length of nucleotide (nt) sequences required for ssi activity; one group that gives rise to larger and clearer plaques and can be reduced to nearly 100 nt (seven out of eleven), and another that generates smaller and less clear plaques and requires more than 200 nt for full activity (four out of eleven). Sequence homology is detected among some members from both groups. The possible biological roles of the ssi are discussed.

Bacteriocin Plasmids↗
PubMed1990

Roles of phi X174 type primosome- and G4 type primase-dependent primings in initiation of lagging and leading strand syntheses of DNA replication.

Eleven single strand initiation sequences (ssi) were isolated from various plasmid genomes using a plaque-morphology assay. Out of seven ssi that require dnaB and dnaC functions for replication in a crude in vitro system, six use a phi X174 type priming mechanism, and a phi X174 type primosome is assembled at these sequences from the purified proteins, n'(priA), n(priB), n"(priC), dnaT, dnaB, dnaC, and primase. The same ssi potentiate dATPase activity of n' protein, and thus represent new n' protein recognition sequences (n'-pas). Based on sequence homology, two structural groups are evident. Two sequences show a strong homology with the phi X174 site, whereas three share extensive homology with the previously characterized n'-pas of ColE1, ssiA(ColE1). All the n'-pas have a potential to form stem and loop structures, although sequence homology between the two classes is absent. In addition to the phi X174 type priming, three ssi do not require either dnaB or dnaC function for replication, and use a G4 type priming, requiring only SSB and primase. The 5' ends of primer RNA synthesized by primase are localized within the vicinity of one of the three blocks of highly conserved nucleotide sequences. Deletions of parts of these conserved sequences result in loss of priming activity, suggesting that they are important for priming on the G4 type ssi, which are termed G site. The general significance of these two types of priming in initiation of lagging or leading strand synthesis as well as various modes of initiation at origins of replication are proposed.

Bacteriophage phi X 174↗
PubMed1990

The ABC-primosome. A novel priming system employing dnaA, dnaB, dnaC, and primase on a hairpin containing a dnaA box sequence.

A priming mechanism requiring dnaA, dnaB, and dnaC proteins operates on a single-stranded DNA coated with single-stranded DNA-binding protein. This novel priming, referred to as "ABC-priming," requires a specific hairpin structure whose stem carries a dnaA protein recognition sequence (dnaA box). In conjunction with primase and DNA polymerase III holoenzyme, ABC-priming can efficiently convert single-stranded DNA into the duplex replicative form. dnaA protein specifically recognizes and binds the single-stranded hairpin and permits the loading of dnaB protein to form a prepriming protein complex containing dnaA and dnaB proteins which can be physically isolated. ABC-priming can replace phi X174 type priming on the lagging strand template of pBR322 in vitro, suggesting a possible function of ABC-priming for the lagging strand synthesis and duplex unwinding. Similar to the phi X174 type priming, a mobile nature of ABC-priming was indicated by helicase activity in the presence of ATP of a prepriming protein complex formed at the hairpin. The implications of this novel priming in initiation of replication at the chromosomal origin, oriC, and in its contribution to the replication fork are discussed.

Bacterial Proteins↗
PubMed1990

The priA gene encoding the primosomal replicative n' protein of Escherichia coli.

The Escherichia coli gene encoding protein n' has been isolated and named priA for primosomal protein A. Protein n' is absolutely required for the conversion of single-stranded phi X174 DNA to the duplex replicative form in an in vitro-reconstituted system. The gene maps to 88.7 minutes on the chromosome adjacent to the cytR locus. Soluble protein extracts from cells harboring the priA gene on a multicopy plasmid contained 45-fold more n' replication activity than wild-type extracts. Enhanced overproduction of greater than 1000-fold was achieved by replacing the natural Shine-Dalgarno sequence with that of the phage T7 phi 10 gene and placing this priA under the control of the T7 phage promoter and RNA polymerase. The priA sequence reveals a 732-amino acid open reading frame and a nucleotide-binding consensus site consistent with the size and ATPase activity of the purified protein. The gene for protein n has been named priB and the putative gene for protein n", priC.

Amino Acid Sequence↗
PubMed1989

Leading strand synthesis of R1 plasmid replication in vitro is primed by primase alone at a specific site downstream of oriR.

By using an in vitro system for R1 plasmid replication dependent on a plasmid-encoded repA protein and host dnaA protein, 5' ends of the nascent leading strand were located at positions 1986-1992, some 380 base pair downstream of oriR. Analyses of early replication intermediates generated in vitro in the presence of dideoxy TTP also indicated that replication initiates about 400 base pair downstream of oriR and proceeds unidirectionally. When a 418-base single-stranded DNA from position 1778 to 2195, derived from the leading strand template, was cloned onto an M13 vector, the chimeric single-stranded phage could be replicated in vitro with only single-stranded DNA binding protein, primase (dnaG gene product), and DNA polymerase III holoenzyme. Furthermore, the priming occurred at a site identical to leading strand initiation. These results strongly suggest that the leading strand synthesis is primed by primase alone. The lagging strand synthesis is specifically terminated at position 1515 or 1516 within oriR, preventing further leftward fork movement. Based on these results, a scheme of R1 plasmid replication is presented.

Bacterial Proteins↗
PubMed1989

Escherichia coli dnaT gene function is required for pBR322 plasmid replication but not for R1 plasmid replication.

Plasmid pBR322 was unable to replicate in a temperature-sensitive dnaT1 strain at a nonpermissive temperature, whereas a pBR322-derived plasmid carrying the wild-type dnaT+ gene was able to replicate under the same conditions. In contrast to pBR322, plasmid R1 could replicate in the dnaT1 strain at a nonpermissive temperature. In keeping with this finding, in vitro replication of plasmid R1 did not require DnaT protein.

DNA Replication↗
PubMed1988

Initiation of lagging-strand synthesis for pBR322 plasmid DNA replication in vitro is dependent on primosomal protein i encoded by dnaT.

The role of the primosome assembly and protein n' recognition site in replication of pBR322 plasmid was examined. The following evidence indicates that the primosome is involved in lagging-strand synthesis of pBR322 plasmid replication in vitro. Early replicative intermediates with newly synthesized leading strand, approximately 1 kilobase pair long, immediately downstream of the replication origin accumulate in products synthesized in extracts from a dnaT strain that lacks primosomal protein i or in wild-type extracts supplemented with anti-protein i antibody. These intermediates are converted efficiently into full-length DNA by addition of purified protein i. Consistent with the previously proposed role of the primosome (Arai, K. and Kornberg, A. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 69-73), an n' site on the lagging strand, but not on the leading strand, is required for efficient replication of the plasmid in vitro. Plasmids lacking an n' site on the lagging strand replicate only to a limited extent in vitro and early replicative intermediates carrying nascent leading strands are accumulated, although a portion of the intermediates complete replication to yield full-length DNA. The latter reaction is completely inhibited by addition of anti-protein i antibody. Insertion of the n' site of phage phi X174 into pBR322 plasmids lacking lagging-strand n' sites restores the replicative ability of the mutant plasmid comparable to that of the wild-type plasmid. These results indicate that protein i is essential for lagging-strand synthesis of pBR322 plasmid in vitro and that it may play an important role in the priming events as a part of either an n' site-dependent primosome or an n' site-independent, as yet unidentified, priming complex.

Bacterial Proteins↗
PubMed1988

Operon structure of dnaT and dnaC genes essential for normal and stable DNA replication of Escherichia coli chromosome.

dnaT and dnaC of Escherichia coli, whose products are components of the primosome, encode genes essential for both normal chromosomal replication and stable DNA replication induced by SOS signals. dnaT and dnaC together with an unknown gene (P-18), which encodes an 18-kDa polypeptide present downstream of dnaC, are cotranscribed from a promoter located 104 base pairs upstream of dnaT. A portion of the transcripts are processed 16 or 27 base pairs upstream of the dnaC coding region and 20% of the transcripts extending through dnaC are terminated at a terminator 103 base pairs downstream of dnaC. The rest of the transcripts pass through this terminator and are terminated downstream of the P-18 gene. The predicted amino acid sequence of the P-18 protein indicate the presence of a 26-amino acid signal peptide at its N terminus, suggesting that this protein may be secreted into the membrane or a periplasmic space. Another gene, which encodes a 14-kDa hydrophobic protein (P-14), was identified upstream of dnaT. The P-14 gene is transcribed from an upstream promoter, and its transcript extends through dnaT and dnaC. These results indicate that dnaT and dnaC constitute an operon with two unknown genes whose products may be associated with a membrane.

Amino Acid Sequence↗
PubMed1988

RepA protein- and oriR-dependent initiation of R1 plasmid replication: identification of a rho-dependent transcription terminator required for cis-action of repA protein.

Initiation of R1 plasmid replication is dependent on cis-acting repA protein and the 188 base-pair (bp) sequence, oriR. RepA protein synthesized in vitro preferentially activates oriR in cis, regardless of the orientation and location of oriR on the template DNA. RepA protein is not reusable after it activates oriR in the cis-position. Cis-action of repA protein is also dependent on the presence of CIS, a 170 bp sequence, between repA and oriR. CIS contains a rho dependent transcription terminator of the repA transcript, deletion of which results in decrease in transformation efficiency and rapid loss of plasmid in the absence of selection. The significance of transcription termination events in replication was indicated by decreased replication activity in vivo caused by premature termination of the repA transcript between repA and CIS. A model which may account for the role of CIS in mediating the cis-action of the repA protein is presented.

Bacterial Proteins↗
PubMed1987

RepA and DnaA proteins are required for initiation of R1 plasmid replication in vitro and interact with the oriR sequence.

RepA, an initiation protein of R1 plasmid replication, was purified from an Escherichia coli strain overproducing the protein. The purified RepA protein specifically initiated replication in vitro of plasmid DNA bearing the replication origin of R1 plasmid (oriR). The replication, strictly dependent on added RepA protein, was independent of host RNA polymerase but required other host replication functions (DnaB and DnaC proteins, the single-stranded-DNA-binding protein SSB, and DNA gyrase). The replication was also completely dependent on the host DnaA function. In filter binding assays in high salt (0.5 M KCl) conditions, RepA specifically binds to both supercoiled and linear plasmid DNA containing the oriR sequence, whereas it binds to nonspecific DNA in low salt. DNase I-protection studies on a linearized DNA fragment revealed that DnaA protein specifically binds to a 9-base-pair DnaA-recognition sequence ("DnaA box") within oriR only when RepA is bound to the sequence immediately downstream of the DnaA box. These results indicate that initiation of R1 plasmid replication is triggered by interaction of RepA and DnaA proteins with the oriR sequence.

Bacterial Proteins↗
PubMed1986

Cloning of the Escherichia coli gene for primosomal protein i: the relationship to dnaT, essential for chromosomal DNA replication.

The Escherichia coli gene encoding one of the primosomal proteins, protein i, was cloned by the use of synthetic oligonucleotide probes. Nucleotide sequence analysis revealed a coding region for protein i of 537 base pairs preceded by a possible promoter sequence. The gene is located adjacent to the dnaC locus, probably both being in a single operon. The protein i gene was shown to be closely related to the dnaT locus based on the following observations. (i) A multicopy plasmid carrying only the protein i gene suppresses the temperature-sensitive phenotype of a dnaT strain and restores the ability of the strain to carry out stable DNA replication in the absence of protein synthesis. (ii) An extract from a dnaT strain does not support replication of the plasmid pBR322 in vitro; addition of purified protein i restores its activity. These results indicate that protein i is encoded by dnaT and that it is essential for chromosomal DNA replication and is involved in the induction of stable DNA replication during the SOS response.

Bacterial Proteins↗
PubMed1983

Cytoarchitecture and visual receptive neurons in the Wulst of the Japanese quail (Coturnix coturnix japonica).

The cellular organization of the Wulst was studied in Nissl- and Golgi-stained brain sections in order to identify the visual receptive neurons. Golgi-impregnated neurons were divided into four types according to their soma size, dendritic configuration, and density of spine distribution. Type I neurons, the largest cells in the Wulst, have long, straight dendrites with many spines. Type II neurons are medium-sized cells with long, straight dendrites. These dendrites have numerous spines. Type III neurons are medium-sized or small cells with spine-free dendrites. Type IV neurons, the smallest cells in the Wulst, have short dendrites with sparse spines. The projections of the nucleus dorsolateralis anterior thalami pars lateralis (DLL) to the Wulst were determined by the Fink-Heimer method. After lesions of the DLL, degenerating terminals are seen in a dorsolateral portion of the nucleus intercalatus hyperstriatum accessorium where the types II, III, and IV neurons are distributed. Postsynaptic elements to the DLL axons were identified by reconstruction of electron microscopic serial sections. Most of the postsynaptic elements were dendritic spines of the type II and IV neurons and a few were dendritic shafts of the type III neurons.

Animals↗
PubMed1983

Ipsilateral retinal projections in Japanese quail, Coturnix coturnix japonica.

Ipsilateral retinal projections were investigated in Japanese quails by means of the Fink-Heimer method after retinal extirpation, and by means of direct injection of horseradish peroxidase or cobalt ion-tophoresis into the optic nerve. Ipsilateral projections were found in the nucleus lateralis anterior thalami, nucleus dorsolateralis anterior thalami pars lateralis, nucleus geniculatus lateralis pars ventralis, nucleus lentiformis mesencephali pars magnocellularis and nucleus ectomamillaris. No ipsilateral retino-tectal projections were observed.

Animals↗
PubMed1983

Definition of oriR, the minimum DNA segment essential for initiation of R1 plasmid replication in vitro.

The 3.6-kilobase Bgl II-EcoRI fragment from R1 plasmid containing copA, repA, and the replication origin (ori) was inserted into the ColE1-type plasmid pUC8. The resulting hybrid plasmid replicates in extracts prepared from both polA- and polA+ cells, whereas pUC8 replicates only in a polA+ extract. This characteristic provides a method for assaying the repA and ori functions. Hybrid plasmids that were either repA- or ori- were unable to replicate in a polA- cell extract. Replication of the repA- ori+ plasmid was restored by complementation of the repA defect by a repA+ ori- plasmid in vitro. Successful complementation of the repA function in vitro provides a method for assaying the repA protein. In order to define the minimum DNA segment with origin function (oriR), deletions were introduced starting from either side of the insert, and the replication properties of the plasmids carrying these deletions were examined in a polA- cell extract. The right end of oriR was located at position 1,611 in the nucleotide coordinates defined previously [Ryder, T., Rosen, J., Armstrong, K., Davidson, D. & Ohtsubo, E. (1981) in The Initiation of DNA Replication: ICN-UCLA Symposia on Molecular and Cellular Biology, ed. Ray, D.S. (Academic, New York), Vol. 22, 91-111]. By complementing repA- ori+ plasmids with the repA+ ori- plasmid, the left end of oriR was localized at position 1,424. Therefore, the oriR sequence, localized within a region of 188 base pairs, is separate from the repA gene. A hybrid plasmid carrying the 206-base-pair segment between positions 1,406 and 1,611 also replicates in a polA- cell extract when the repA function is supplied in trans. Removal of an additional 66 base pairs (positions 1,406-1,471) inactivates the function of the minimal oriR segment.

Base Sequence↗
PubMed1983

Brain organization of sharks, with special reference to archaic species.

Using photographic measurements and normal neurohistological preparations, a number of brains of Galeomorph and Squalomorph sharks were studied and compared. The brains are generally more developed in Galeomorph than in Squalomorph sharks. Among the major brain territories in Galeomorph sharks, the nucleus centralis is prominently enlarged, and the corpus cerebelli is much swollen and fissured. In archaic species such as Scapanorhynchus, Heterodontus, and Chlamydoselachus, the cerebral pallium and corpus cerebelli are less developed than those of taxonomically close species. These results were discussed, in relation to taxonomy, phylogeny and ecology.

Animals↗
PubMed1982

Decreased synaptic areas on Purkinje spines in the cerebellar cortex of the mutant Japanese quail, Coturnix coturnix japonica.

'Dark frayed feather nervous disorder' (dn) is a neurological mutation in quails in which the cerebellar cortex is abnormally organized. Purkinje cells are not aligned in a single row and show hypoplasia of the dendrites. The synapses between the parallel fibers and the spines of the Purkinje cell dendrites were examined with the technique of serial sections in electron microscopy. The postsynaptic thickenings were obviously decreased in the mutant quail despite the same density and size of dendritic spines of Purkinje cells. In addition, ectopic spines and postsynaptic differentiations free of parallel fibers were not found on the dn Purkinje cell. Because of the poor dendritic arborization, the total number of spines and the total synaptic area are, therefore, reduced in the dn Purkinje cell. According to the results obtained the dn mutant genetic locus is considered to affect primarily Purkinje cells.

Animals↗