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A Fratini

Publications and source records attributed to A Fratini.

14 recordsLinked to original sources

Dietary cysteine regulates the levels of mRNAs encoding a family of cysteine-rich proteins of wool.

The abomasal or intravenous infusion of sulphur-containing amino acids such as cysteine or methionine into sheep on low-quality diets increases the sulphur content of the wool by increasing the synthesis of proteins containing a cysteine content of approximately 30 mol %. To investigate the molecular and cellular basis of this nutritional effect, quantitative analyses of wool keratin mRNA and protein levels, and follicle cortical cell type, were undertaken in sheep intravenously infused with cysteine. Northern blot analyses revealed that the mRNA levels of one gene family encoding cysteine-rich keratin-associated proteins (KAP4 family) expressed in the wool follicle cortex, increased approximately 5-6 times. Furthermore, the response was rapid as the mRNA levels increased approximately 3.5 times after 1 d of the cysteine infusion and, by 1 d post-infusion, they had fallen, approaching their basal level. No changes in the mRNA levels encoding the intermediate filament or the other keratin-associated protein families of lower cysteine content were observed. Concomitantly, two-dimensional polyacrylamide gel electrophoresis analysis of wool proteins showed a striking increase in the abundance of a group of cysteine-rich keratin-associated proteins in the wool by the end of the infusion period, returning to basal levels by 3 weeks later. At the cellular level, KAP4 expression was localized to the follicle paracortical cells, and the proportion of paracortical cells and the extent of KAP4 expression paralleled the changes in the cysteine infusion status of the sheep.

Amino Acid Sequence↗

Sequence, expression, and evolutionary conservation of a gene encoding a glycine/tyrosine-rich keratin-associated protein of hair.

In hair differentiation several families of keratin proteins with distinctive amino acid compositions are produced. To study the role and regulation of one of these families, the glycine/tyrosine-rich keratin-associated proteins encoded by the KAP6 gene family, a partial wool follicle cDNA clone encoding a sheep KAP6 protein was sequenced and the corresponding gene isolated from a sheep cosmid library. The KAP6.1 gene encodes a basic protein of 82 amino acids (M(r) = 8,296) with a combined glycine and tyrosine content of approximately 60 mol%. There are several KAP6 genes in the sheep genome, all located within a 1,050-kilobase SfiI fragment. Northern blot analysis demonstrated that at least one member of the KAP6 family is expressed in the wool follicle. A rabbit KAP6 gene was isolated and its sequence and expression patterns were compared with the sheep gene. The sheep and rabbit genes have a nucleotide sequence identity of 89%, suggesting that they are equivalent genes and indicating strong selection pressure during evolution. Both genes contain several conserved sequence motifs of 7-9 nucleotides in their 5'-flanking regions that may be involved in the regulation of their expression. Localization of KAP6 mRNAs in sheep wool and rabbit hair follicles by in situ hybridization suggests that the genes are expressed in the cells of the hair shaft cortex in varying expression patterns. KAP6 expression starts relatively late in hair follicle differentiation, and the proportion of hair cortical cells that express it may change from follicle to follicle.

Amino Acid Sequence↗

A linkage group with FRA16B (the fragile site at 16q22.1).

Polymorphic DNA markers located in bands 16q13, 16q21 and 16q22 were examined for recombination with FRA16B, the fragile site at 16q22.100. A tight linkage cluster D16S10-FRA16B-D16S4-HP was established. There were no recombinants (theta = 0.0, z = 8.3) between D16S10 and D16S4, which flank FRA16B. The markers D16S10 and D16S4 are in close proximity on the genetic map and delineate a small chromosomal segment, which contains the distamycin A-inducible fragile site.

Chromosome Fragile Sites↗

Mapping the short arm of human chromosome 16.

Physical mapping of 13 different breakpoints on the short arm of chromosome 16 using previously mapped probes and the subsequent mapping of additional probes enabled the division of this portion of the chromosome into six different intervals. D16S94 was mapped between HBA and D16S80 and is closer to PKD1 than either HBA or D16S80. A tight linkage group which includes FRA16A, D16S8, and D16S79 was identified. Seven breakpoints, including FRA16A, could not be separated by probe localizations. This study provides the basis for the development of detailed maps of the short arm of chromosome 16.

Animals↗

Translocation breakpoint in t(11;14) in B-cell leukemia is not at the rare fragile site at 11q13.3.

The cloned breakpoint at 11q13.3 of the t(11;14)(q13.3;q32.3) in a B-cell lymphocytic leukemia (B-CLL) was used to analyze DNA from individuals with and without the rare folate-sensitive fragile site at 11q13.3. On Southern blots there were no discernible differences. Subclones of the ends of the leukemia breakpoint clone were prepared and used for in situ hybridization to chromosomes expressing fra(11)(q13.3). Both subclones hybridized distal to the fragile site. These experiments indicate that the breakpoints at 11q13.3 in B-CLL (and in a B-cell lymphoma) are not at the fragile site at 11q13.3.

B-Lymphocytes↗

Fine mapping of gene probes and anonymous DNA fragments to the long arm of chromosome 16.

The fragile site, FRA16B, at 16q22.100 and four different translocations with breakpoints at 16q22.102, 16q22.105, 16q22.108, and 16q22.3 were used to locate and order DNA probes. This was achieved by Southern analysis of a somatic cell hybrid panel containing portions of chromosome 16 and by in situ hybridization. The anonymous DNA fragments D16S6, D16S10, and D16S11 were proximal to FRA16B and located at 16q13----q22.100. D16S4 and LCAT were located at 16q22.100----q22.102. TAT and HP were located at 16q22.105----q22.108. CTRB was located distal to 16q22.105 and therefore is in the distal half of 16q22. The order of markers in this region was determined as centromere-D16S6, D16S11, D16S10, MT-FRA16B-D16S4, LCAT-HP,TAT,CTRB-APRT- telomere. Linkage studies to determine map distances between the closest markers flanking the fragile site are now in progress.

Animals↗

Molecular genetics of human chromosome 16.

The major diseases mapped to chromosome 16 are adult polycystic kidney disease and those resulting from mutations in the alpha globin complex. There are at least six other less important genetic diseases which map to this chromosome. The adenine phosphoribosyltransferase gene allows for selection of chromosome 16 in somatic cell hybrids and a hybrid panel is available which segments the chromosome into six regions to facilitate gene mapping. Genes which have been mapped to this chromosome or which have had their location redefined since HGM8 include APRT, TAT, MT, HBA, PKD1, CTRB, PGP, HAGH, HP, PKCB, and at least 19 cloned DNA sequences. There are RFLPs at 13 loci which have been regionally mapped and can be used for linkage studies.

Adenine Phosphoribosyltransferase↗

A new location for the human adenine phosphoribosyltransferase gene (APRT) distal to the haptoglobin (HP) and fra(16)(q23)(FRA16D) loci.

The human adenine phosphoribosyltransferase gene (APRT) was mapped with respect to the haptoglobin gene (HP) and the fragile site at 16q23.2 (FRA16D). A subclone of APRT and a cDNA clone of HP were used for molecular hybridization to DNA from mouse-human hybrid cell lines containing specific chromosome 16 translocations. The APRT subclone was used for in situ hybridization to chromosomes expressing FRA16D. APRT was found to be distal to HP and FRA16D and was localized at 16q24, making the gene order cen-FRA16B-HP-FRA16D-APRT-qter.

Adenine Phosphoribosyltransferase↗

Excess thymidine induces folate sensitive fragile sites.

Folate sensitive fragile sites on human chromosomes have been found to be inducible in cultured lymphocytes by high levels of thymidine but not by high levels of BrdU. The biochemical interpretation of events leading to fragile site expression has been revised since it is now clear that low levels of either thymidylate or deoxycytidine triphosphate will result in this phenomenon. A model for the DNA at a fragile site, composed of alternating repeating polypurine/polypyrimidine sequences is proposed.

Base Sequence↗