Mismatching and the Reassociation Rate of Mouse Satellite DNA

Sutton, W.D.; McCallum, Mora

doi:10.1038/newbio232083a0

Cited by 52 publications

(20 citation statements)

References 14 publications

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“…Furthermore, the ATm of the high-melting fraction remains constant with varying conditions of stringency during reassociation (legend, Table 1). These findings indicate that the fidelity of base pairing in the reformed duplexes is almost perfect and that reassociation kinetics may be interpreted without considering the effect of mispairing on reassociation rate (18)(19)(20).…”

Section: Resultsmentioning

confidence: 89%

“…About one-third of the muskmelon satellite is high melting and is a simple repeating sequence (about 380 np). This is the approximate complexity of all animal satellites thus far reported (7,19,40). The remaining two-thirds of the muskmelon satellite is low melting, is not covalently linked with the simple sequence, and is about 103-fold greater in complexity than the simple sequence.…”

Section: Discussionmentioning

confidence: 95%

“…The remaining two-thirds of the muskmelon satellite is low melting, is not covalently linked with the simple sequence, and is about 103-fold greater in complexity than the simple sequence. Furthermore, the components of the satellite are composed of essentially perfect multiple copies, rather than the less precisely "repeated" sequences (ATm of 3-23°) found in most animal satellites (7,10,19,21,40,41). Satellite DNA of Drosophila virilis has a ATm of 2° (16).…”

Section: Discussionmentioning

confidence: 99%

“…Sequencing techniques have also provided length estimates of ten in a kangaroo rat satellite (16) and seven for each of three satellites in Drosophila virilis (17). The original complexity estimate (2) for mouse satellite of 300-400 nucleotide pairs (np) was considered too large after estimates were made of the marked influence of mispairing on reassociation rate (18,19 …”

Section: Introductionmentioning

confidence: 99%

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Novel Properties of Satellite DNA from Muskmelon

Bendich¹,

Anderson²

1974

Proc. Natl. Acad. Sci. U.S.A.

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The purified dense satellite of muskmelon (Cucumis melo L.) represents 30% of the total DNA and forms a sharp, unimodal peak in both neutral and alkaline CsCl gradients. Spectrophotometric melting and DNA reassociation analyses revealed that one-third of the satellite is high melting and has a complexity of about 2.5 X 105 daltons, while the remaining two-thirds of the satellite melts 80 lower and has a complexity of about 5 X 108 daltons. The thermal stability of reassociated satellite DNA indicates that the multiple copies of the two melting fractions are essentially identical. The sharp, unimodal peak in Cs2SO4 becomes two distinct peaks when either silver or mercuric ions are included in the Cs2 S04 gradient One or more extremely rapidly reassociating components of nuclear DNA are present in many animals, often detectable as "satellite" bands in CsCl gradients (1). The kinetics of strand reassociation and the stability of these reformed duplexes are interpreted to mean that the multiple DNA segments are highly reiterated and are closely similar copies (2-4). Although its function is not known, some satellite DNA in mice (5) and insects (6, 7) hybridizes primarily with the centromeric regions of chromosomes in cytological preparations. It is enriched in heterochromatin of guinea pig (8), mouse, crab (9) and calf (10), and in mouse nucleoli (11). The mouse satellite does not appear to be transcribed (12). These unusual properties have led to the suggestion that satellite DNA may have a structural function in chromosome folding, pairing, and movement (12,13).From its reassociation kinetics, mouse satellite was estimated to contain about a million very similar copies comprising some 10% of the total mouse DNA (2, 14). The repeating unit in guinea pig a-satellite DNA has been estimated from oligonucleotide digestion products to be six nucleotides long (somewhat longer in the mouse) with about 12% nucleotide changes in the repeating units (15). Sequencing techniques have also provided length estimates of ten in a kangaroo rat satellite (16) and seven for each of three satellites in Drosophila virilis (17). The original complexity estimate (2) for mouse satellite of 300-400 nucleotide pairs (np) was considered too large after estimates were made of the marked influence of mispairing on reassociation rate (18,19

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Section: Resultsmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Novel Properties of Satellite DNA from Muskmelon

Bendich¹,

Anderson²

1974

Proc. Natl. Acad. Sci. U.S.A.

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“…Decreased annealing rate can be due to the formation of poorly matched hybrids (20), yet the hybrids formed here were well-matched. They had a high Tm and sharp melting curve like M-321 mRNA hybrids (Fig.…”

Section: Methodsmentioning

confidence: 99%

Region of immunoglobulin light-chain mRNA transcribed into complementary DNA by RNA-dependent DNA polymerase of avian myeloblastosis virus.

Schechter¹

1975

Proc. Natl. Acad. Sci. U.S.A.

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The mRNA coding for a K-type immunoglobulin light (L)chain and its complementary DNA (cDNA) hybridize with a CrtI/2 of 2.6 X 10-4 moles of ribonucleotide X liter-l X sec, forming well-matched duplexes (melting temperature Tm = 890). The molecular weight of the cDNA is about 280,000 (840 nucleotides) as determined by alkaline sucrose gradient centrifugation and from the extent of protection ofthe mRNA by the cDNA from ribonuclease digestion. The distribution of genetic markers on immunoglobulin (Ig) chains (1) and amino-acid sequence data (2) raised the possibility that different genes code for the variable (V, N-terminal) and constant (C, C-terminal) parts of the intact polypeptide chain. According to the somatic mutation hypothesis, the genome contains a few V-region genes, whereas the germ line hypothesis predicts a large number of V-region genes. To resolve this issue requires highly labeled nucleic acid probes that are specific to V-and C-region sequences. Furthermore, it is necessary to find out to what extent a probe of one V-region would detect and quantify nucleic acid sequences of other V-regions. It is possible that a given V-region probe would anneal with V-regions coding for light (L)-chains of the same subgroup (i.e., L-chains having similar amino-acid sequence at the V-region) but would not detect V-regions of L-chains belonging to different subgroups.The isolation of Ig L-chain mRNA from myeloma polysomes specifically precipitated with antibodies to L-chain has been achieved. Over 95% of the cell-free product programmed by the mRNA was L-chain protein, as determined from two-dimensional tryptic fingerprints (3, 4). We here report the enzymic synthesis and characterization of the DNA complementary to this highly purified mRNA.Abbreviations: AMV, avian myeloblastosis virus; cDNA, complementary DNA; 1g, immunoglobulin; L-chain, light chain; V-region, variable region of immunoglobulin; C-region, constant region; Crt, product of RNA concentration (moles of nucleotide per liter) and time (sec); Tm, melting temperature. MATERIALS AND METHODSThe mRNAs coding for M-321, M-63, M-41, and M-104E L-chains were prepared from the respective mouse myeloma polysomes specifically precipitated with antibodies to L-chains, followed by chromatography on oligo(dT)-cellulose (3, 4). Over 95% of the cell-free products programmed in vitro (3) by each mRNA corresponded to the appropriate L-chain. When analyzed on 3.2% polyacrylamide gels made in 98% formamide (5), all mRNAs migrated as 15.5 + 0.5S bands, and they contained variable amounts of rRNA (Table 1). The M-321 mRNA was further purified by sucrose gradient centrifugation to yield preparation M-321p, having only 5% rRNA contamination. The DNA complementary to M321p mRNA (cDNA) was synthesized with the aid of the avian myeloblastosis virus (AMV) DNA polymerase (6) in the presence of [3H]dCTP (26 Ci/mmol), actinomycin D (0.01%), and (dT)10 primer. Hybridizations were performed at 68°in 0.02 M Na-phosphate (pH 7.0), 0.3 M NaCl, 2 mM EDTA, and 0.1% sodium dodecy...

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The effect of thymine dimers on DNA: DNA hybridization

Kahn

1974

Biopolymers

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synopsisDNA from bacteriophage T7 was irradiated at long ultraviolet wavelengths in the presence of silver ions. Such treatment leads to selective production of thymine: thymine dimen in DNA. The DNA was melted and the renaturation rate was determined as a function of thymine dimer content and renaturation temperature. Under "normal" hybIidieation conditions little change in the renaturation rate was observed even when 30% of the thymine was dimerised. This result is consistent with the view that up to a 15% change in the primary sequence of DNA does not appreciably change the renaturation rate.The kinetics of nucleic acid hybridization have been shown to be dependent on a variety of factors such as temperature, ionic strength, and guanine-cytosine c0ntent.l A long-standing question is to what extent a mismatch of bases (i.e., unconventional pairing of bases) will affect the renaturation rate.' Since the measurable parameter in the Wetmur and Davidson formulation2 of renaturation kinetics K , contains both a complexity correction factor and a putative mismatch correction factor, the estimation of complexity based solely on K N might be questioned in cases in which mismatch is known to be p r e~e n t .~.~ As an example of the effect of mismatching, reannealed hybrids of the rapidly renaturing fraction of eukaryotic DNA show a decreased melting temperature after renaturation, indicating that the hybrid contains mismatched bases.5 Direct sequence analysis of the guinea pig a satellite DNA suggests that mutational change in the primary DNA sequence is common and will lead to mismatch upon hybridization. Sutton and RlcCallum4 reannealed mouse satellite DNA and subsequently separated the duplexes into four classes on the basis of differing melting temperature

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Mismatching and the Reassociation Rate of Mouse Satellite DNA

Cited by 52 publications

References 14 publications

Novel Properties of Satellite DNA from Muskmelon

Novel Properties of Satellite DNA from Muskmelon

Region of immunoglobulin light-chain mRNA transcribed into complementary DNA by RNA-dependent DNA polymerase of avian myeloblastosis virus.

The effect of thymine dimers on DNA: DNA hybridization

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