Identification of mouse SLC39A8 as the transporter responsible for cadmium-induced toxicity in the testis

Dalton et al. 10.1073/pnas.0406085102.

Supporting Information

Files in this Data Supplement:

Supporting Table 1
Supporting Table 2
Supporting Table 3
Supporting Figure 5

Fig. 5. DNA sequence upstream of exon 2 of the mouse Slc39a8 gene, showing the three alternative first exons (bold). Translation of the ZRT-, IRT-like protein (ZIP)8 protein begins at +233 of exon 2; the initiation codon ATG is underlined. Note that all three introns after the alternative exons 1 have acceptor sites GT that can join with the donor site AG immediately before exon 2. The remaining portions of exon 2 and exons 3–9 are not shown. The human SLC39A8 gene likewise has three exons 1 (data not shown).

Table 1. Primers used in progeny from the B6D2F₁´ B6 backcross

Markers	Primer F (sequence)	Primer R (sequence)
D3Mit110	AGCCCTAGGATTTGCATGG	TTGGGGTTTTGACTTTTATTTATT
M15-2	AGGTGCCAGAGCCAAAAA	GACAGGGGAAACAGAACTATCC
M13-2	TCATGTTGTATTTATGCACTTAGGA	ACTTTGGACATAGGGAAATTCTG
M12-17	GGCTTAAAAGAGAGTACCGGG	TTCCACCACTTCCCTTTACAA
M12-7	CAAAATGACATAAAACTATCACATGC	CATTTCAAAACATCATCCATAAAA
M11-22	AAGTGCATCTAAAGAAAGGAAATG	CACTTAACAAGTCTTTGAAAATGGA
M11-6	TTTATTTCCTTTGAGATGAGGG	TGATAAGATGACAATCAATGCTG
M49-18	ATTATTGGTGAGGGGCAGAG	AATCCATGCATGTGGACGTA
S530	AGGGCTGTGCACCTTTCTTA	TGGCACTACACTCCCTTTCC
S901	CCCTTGCTGATGAAGAGACC	CCAGCTCTGCCTAAGCTTGT
S101	TCTCCTCTAATGGGCTGGAA	TGGTTTACTACCGAGTGAGCA
S131	CAGCTCAACAGGAGGGAAAC	AAAATCTTGCCACGTGCTCT
S132	GTTGGGTGGGAAAACAACAG	GGGAACAACCTAAGGGCAAT
M49-11	GGGGTTGCATCTGTCTCTTT	TCATGCACACACACATGGAC
M9-9	TGAATTTCCACTTACAGTTCACTTG	TTTCTTCAGTTTCTATAATAACCCCC
D3Mit291	AACTTCATGTACTCTCTTTCTCTCAGC	CCACAAGTAGGACAACGCAA
M10-26	TCTAGAGGAACAGAACAGAAGGA	TCAGTTTCTAAGGCAATCTAGTGA
D3Mit255	TGACACAAACTTGCATTATCTGG	TTACTTAAGAAACTTGCCCTCCC

The positions of these markers on mouse chromosome 3 can be seen in Fig. 1c. Primers: F, forward; R, reverse.

Table 2. Positions of the informative SNPs when analyzing progeny from the B6D2F₁´ B6 backcross

SNP sequence	SNP sequence
S530	GAGCACTGTG(A/G)GGAGGGAGCG
S901	GTCACACAAA(A/G)CGAAGGAAAG
S101	GAGTACAGCA(A/T)CTGGTGACAT
S131	GTCCTCACCC(T/C)TCTGCCAGGA
S132	ACCTCAATAC(T/C)GGACACAAGA

The bolded biallelic nucleotide site [e.g., (A/G)] in the middle of each primer denotes the informative SNP that helped in identifying the nine new recombinant offspring, which are listed in Fig. 1c.

Table 3. Comparison of the intron-exon structure of the mouse and human SLC39A8 genes

	Exon, bp		Intron, bp
No.	Mouse	Human	Mouse	Human
1c	256	184*	708	801*
1b	144	139*	439	429*
1a	37	26*	156	159*
2	451	696	26,096	28,615
3	163	163	6,812	8,064
4	170	170	1,410	2,326
5	123	123	467	509
6	171	165	25,968	36,239
7	208	208	199	199
8	185	185	2,020	4,298
9	1,587^†	1,530^†
Total (excluding exons/introns 1)	3,058^‡	3,240^‡	62,972	80,250

*Human exons and introns 1c, 1b, and 1a are theoretical but extracted by human/mouse/rat comparative genomics alignments; the exons and introns match the mouse and rat Slc39a8 sequences sufficiently but are not yet supported by ESTs in the human database.

^†Longest possible exon 9 out of many transcripts in the EST database.

^‡Longest possible transcript though numerous shorter ones are found in the EST database.