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Species-Specific Expression of Full-Length and Alternatively Spliced Variant Forms of CDK5RAP2.

Park JS, Lee MK, Kang S, Jin Y, Fu S, Rosales JL, Lee KY - PLoS ONE (2015)

Bottom Line: CDK5RAP2 is one of the primary microcephaly genes that are associated with reduced brain size and mental retardation.We have previously shown that human CDK5RAP2 exists as a full-length form (hCDK5RAP2) or an alternatively spliced variant form (hCDK5RAP2-V1) that is lacking exon 32.Here, we demonstrate that rat expresses both a full length and an alternatively spliced variant form of CDK5RAP2 that are equivalent to our previously reported hCDK5RAP2 and hCDK5RAP2-V1, repectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Anatomy, Southern Alberta Cancer Research and Hotchkiss Brain Institutes, University of Calgary, Calgary, Alberta, Canada.

ABSTRACT
CDK5RAP2 is one of the primary microcephaly genes that are associated with reduced brain size and mental retardation. We have previously shown that human CDK5RAP2 exists as a full-length form (hCDK5RAP2) or an alternatively spliced variant form (hCDK5RAP2-V1) that is lacking exon 32. The equivalent of hCDK5RAP2-V1 has been reported in rat and mouse but the presence of full-length equivalent hCDK5RAP2 in rat and mouse has not been examined. Here, we demonstrate that rat expresses both a full length and an alternatively spliced variant form of CDK5RAP2 that are equivalent to our previously reported hCDK5RAP2 and hCDK5RAP2-V1, repectively. However, mouse expresses only one form of CDK5RAP2 that is equivalent to the human and rat alternatively spliced variant forms. Knowledge of this expression of different forms of CDK5RAP2 in human, rat and mouse is essential in selecting the appropriate model for studies of CDK5RAP2 and primary microcephaly but our findings further indicate the evolutionary divergence of mouse from the human and rat species.

No MeSH data available.


Related in: MedlinePlus

A. The equivalent of exon 32 in full-length rat cdk5RAP2 (rcdk5RAP2) is absent in the alternatively spliced variant form of rat cdk5RAP2 (rcdk5RAP2-v1) and in the mouse full-length cdk5RAP2 (mCDk5rap2).Sequences of rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are aligned and amino acid numbers are indicated on either side. The identical sequences in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are shown. The broken lines correspond to the amino acid sequence missing in rcdk5RAP2-v1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1. The codon (GAG) encoding glutamic acid (E, shaded in grey) in rCDK5RAP2 and hCDK5RAP2 is generated from the combination of nucleotides (G-AG) between exon 31 and exon 32. The codon (GAT) encoding aspartic acid (D, shaded in grey) in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 is generated from the combination of nucleotides (G-AT) between exon 32 and exon 33 in rCDK5RAP2 and hCDK5RAP2, and exon 31 and exon 32 in rCDK5RAP2-V1, mCDK5RAP2 and hCDK5RAP2-V1, respectively. B. Primary structures of rCDK5RAP2, rCDK5RAP2-V1 and mCDK5RAP2. The amino acid numbers for the respective domains were based on sequence alignment with hCDK5RAP2 using CLUSTAL 2.1 multiple sequence alignment software. rCDK5RAP2 encodes 1903 amino acids while rCDK5RAP2-V1 encodes 1820 amino acids lacking 83 amino acids (aa1582-aa1664) that are found in rCDK5RAP2. The missing amino acids (1582 to 1664) in rCDK5RAP2-V1 are also absent in mCDK5RAP2, which encodes 1822 amino acids. Note that the C-terminal portion of the second SMC domain in rCDK5RAP2 is absent in rCDK5RAP2-V1 and mCDK5RAP2 as well. hCDK5RAP2 encodes 1,893 amino acids (215 kDa) while hCDK5RAP2-V1 encodes 1,814 amino acids (206 kDa) lacking 79 amino acids from amino acid1,576 to amino acid1,654. hCDK5RAP2 (genbank accession no.NP_060719) and hCDK5RAP2-V1 (genbank accession no. NP_001011649) are shown.
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pone.0142577.g003: A. The equivalent of exon 32 in full-length rat cdk5RAP2 (rcdk5RAP2) is absent in the alternatively spliced variant form of rat cdk5RAP2 (rcdk5RAP2-v1) and in the mouse full-length cdk5RAP2 (mCDk5rap2).Sequences of rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are aligned and amino acid numbers are indicated on either side. The identical sequences in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are shown. The broken lines correspond to the amino acid sequence missing in rcdk5RAP2-v1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1. The codon (GAG) encoding glutamic acid (E, shaded in grey) in rCDK5RAP2 and hCDK5RAP2 is generated from the combination of nucleotides (G-AG) between exon 31 and exon 32. The codon (GAT) encoding aspartic acid (D, shaded in grey) in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 is generated from the combination of nucleotides (G-AT) between exon 32 and exon 33 in rCDK5RAP2 and hCDK5RAP2, and exon 31 and exon 32 in rCDK5RAP2-V1, mCDK5RAP2 and hCDK5RAP2-V1, respectively. B. Primary structures of rCDK5RAP2, rCDK5RAP2-V1 and mCDK5RAP2. The amino acid numbers for the respective domains were based on sequence alignment with hCDK5RAP2 using CLUSTAL 2.1 multiple sequence alignment software. rCDK5RAP2 encodes 1903 amino acids while rCDK5RAP2-V1 encodes 1820 amino acids lacking 83 amino acids (aa1582-aa1664) that are found in rCDK5RAP2. The missing amino acids (1582 to 1664) in rCDK5RAP2-V1 are also absent in mCDK5RAP2, which encodes 1822 amino acids. Note that the C-terminal portion of the second SMC domain in rCDK5RAP2 is absent in rCDK5RAP2-V1 and mCDK5RAP2 as well. hCDK5RAP2 encodes 1,893 amino acids (215 kDa) while hCDK5RAP2-V1 encodes 1,814 amino acids (206 kDa) lacking 79 amino acids from amino acid1,576 to amino acid1,654. hCDK5RAP2 (genbank accession no.NP_060719) and hCDK5RAP2-V1 (genbank accession no. NP_001011649) are shown.

Mentions: As shown in Fig 3A and 3B, sequencing analysis of the 352 and 103 bp PCR products and subsequent analysis of deduced amino acid sequence confirmed that the full-length rCDK5RAP2 (GenBank, accession number: JX524852) contains exon 32 and encodes 1903 amino acids (215 kDa).


Species-Specific Expression of Full-Length and Alternatively Spliced Variant Forms of CDK5RAP2.

Park JS, Lee MK, Kang S, Jin Y, Fu S, Rosales JL, Lee KY - PLoS ONE (2015)

A. The equivalent of exon 32 in full-length rat cdk5RAP2 (rcdk5RAP2) is absent in the alternatively spliced variant form of rat cdk5RAP2 (rcdk5RAP2-v1) and in the mouse full-length cdk5RAP2 (mCDk5rap2).Sequences of rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are aligned and amino acid numbers are indicated on either side. The identical sequences in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are shown. The broken lines correspond to the amino acid sequence missing in rcdk5RAP2-v1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1. The codon (GAG) encoding glutamic acid (E, shaded in grey) in rCDK5RAP2 and hCDK5RAP2 is generated from the combination of nucleotides (G-AG) between exon 31 and exon 32. The codon (GAT) encoding aspartic acid (D, shaded in grey) in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 is generated from the combination of nucleotides (G-AT) between exon 32 and exon 33 in rCDK5RAP2 and hCDK5RAP2, and exon 31 and exon 32 in rCDK5RAP2-V1, mCDK5RAP2 and hCDK5RAP2-V1, respectively. B. Primary structures of rCDK5RAP2, rCDK5RAP2-V1 and mCDK5RAP2. The amino acid numbers for the respective domains were based on sequence alignment with hCDK5RAP2 using CLUSTAL 2.1 multiple sequence alignment software. rCDK5RAP2 encodes 1903 amino acids while rCDK5RAP2-V1 encodes 1820 amino acids lacking 83 amino acids (aa1582-aa1664) that are found in rCDK5RAP2. The missing amino acids (1582 to 1664) in rCDK5RAP2-V1 are also absent in mCDK5RAP2, which encodes 1822 amino acids. Note that the C-terminal portion of the second SMC domain in rCDK5RAP2 is absent in rCDK5RAP2-V1 and mCDK5RAP2 as well. hCDK5RAP2 encodes 1,893 amino acids (215 kDa) while hCDK5RAP2-V1 encodes 1,814 amino acids (206 kDa) lacking 79 amino acids from amino acid1,576 to amino acid1,654. hCDK5RAP2 (genbank accession no.NP_060719) and hCDK5RAP2-V1 (genbank accession no. NP_001011649) are shown.
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pone.0142577.g003: A. The equivalent of exon 32 in full-length rat cdk5RAP2 (rcdk5RAP2) is absent in the alternatively spliced variant form of rat cdk5RAP2 (rcdk5RAP2-v1) and in the mouse full-length cdk5RAP2 (mCDk5rap2).Sequences of rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are aligned and amino acid numbers are indicated on either side. The identical sequences in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 are shown. The broken lines correspond to the amino acid sequence missing in rcdk5RAP2-v1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1. The codon (GAG) encoding glutamic acid (E, shaded in grey) in rCDK5RAP2 and hCDK5RAP2 is generated from the combination of nucleotides (G-AG) between exon 31 and exon 32. The codon (GAT) encoding aspartic acid (D, shaded in grey) in rCDK5RAP2, rCDK5RAP2-V1, mCDK5RAP2, hCDK5RAP2 and hCDK5RAP2-V1 is generated from the combination of nucleotides (G-AT) between exon 32 and exon 33 in rCDK5RAP2 and hCDK5RAP2, and exon 31 and exon 32 in rCDK5RAP2-V1, mCDK5RAP2 and hCDK5RAP2-V1, respectively. B. Primary structures of rCDK5RAP2, rCDK5RAP2-V1 and mCDK5RAP2. The amino acid numbers for the respective domains were based on sequence alignment with hCDK5RAP2 using CLUSTAL 2.1 multiple sequence alignment software. rCDK5RAP2 encodes 1903 amino acids while rCDK5RAP2-V1 encodes 1820 amino acids lacking 83 amino acids (aa1582-aa1664) that are found in rCDK5RAP2. The missing amino acids (1582 to 1664) in rCDK5RAP2-V1 are also absent in mCDK5RAP2, which encodes 1822 amino acids. Note that the C-terminal portion of the second SMC domain in rCDK5RAP2 is absent in rCDK5RAP2-V1 and mCDK5RAP2 as well. hCDK5RAP2 encodes 1,893 amino acids (215 kDa) while hCDK5RAP2-V1 encodes 1,814 amino acids (206 kDa) lacking 79 amino acids from amino acid1,576 to amino acid1,654. hCDK5RAP2 (genbank accession no.NP_060719) and hCDK5RAP2-V1 (genbank accession no. NP_001011649) are shown.
Mentions: As shown in Fig 3A and 3B, sequencing analysis of the 352 and 103 bp PCR products and subsequent analysis of deduced amino acid sequence confirmed that the full-length rCDK5RAP2 (GenBank, accession number: JX524852) contains exon 32 and encodes 1903 amino acids (215 kDa).

Bottom Line: CDK5RAP2 is one of the primary microcephaly genes that are associated with reduced brain size and mental retardation.We have previously shown that human CDK5RAP2 exists as a full-length form (hCDK5RAP2) or an alternatively spliced variant form (hCDK5RAP2-V1) that is lacking exon 32.Here, we demonstrate that rat expresses both a full length and an alternatively spliced variant form of CDK5RAP2 that are equivalent to our previously reported hCDK5RAP2 and hCDK5RAP2-V1, repectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Anatomy, Southern Alberta Cancer Research and Hotchkiss Brain Institutes, University of Calgary, Calgary, Alberta, Canada.

ABSTRACT
CDK5RAP2 is one of the primary microcephaly genes that are associated with reduced brain size and mental retardation. We have previously shown that human CDK5RAP2 exists as a full-length form (hCDK5RAP2) or an alternatively spliced variant form (hCDK5RAP2-V1) that is lacking exon 32. The equivalent of hCDK5RAP2-V1 has been reported in rat and mouse but the presence of full-length equivalent hCDK5RAP2 in rat and mouse has not been examined. Here, we demonstrate that rat expresses both a full length and an alternatively spliced variant form of CDK5RAP2 that are equivalent to our previously reported hCDK5RAP2 and hCDK5RAP2-V1, repectively. However, mouse expresses only one form of CDK5RAP2 that is equivalent to the human and rat alternatively spliced variant forms. Knowledge of this expression of different forms of CDK5RAP2 in human, rat and mouse is essential in selecting the appropriate model for studies of CDK5RAP2 and primary microcephaly but our findings further indicate the evolutionary divergence of mouse from the human and rat species.

No MeSH data available.


Related in: MedlinePlus